Kinase antagonists

ABSTRACT

The present invention provides novel compounds that are antagonists of PI3 kinase, PI3 kinase and tryosine kinase, PI3Kinase and mTOR, or PI3Kinase, mTOR and tryosine kinase.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 60/744,269, filed Apr. 4, 2006, and U.S. ProvisionalPatent Application No. 60/744,270, filed Apr. 4, 2006, both of which areincorporated herein by reference in their entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

The present invention was supported by a grant from the NationalInstitutes of Health (AI44009). The Government has certain rights to theinvention.

BACKGROUND OF THE INVENTION

Phosphoinositide 3-kinases (PI3-Ks) catalyze the synthesis of thephosphatidylinositol (PI) second messengers PI(3)P, PI(3,4)P2; andPI(3,4,5)P3 (PIP3) (Fruman et al., 1998). In the appropriate cellularcontext, these three lipids control diverse physiological processesincluding cell growth, survival, differentiation and chemotaxis (Katsoet al., 2001). The PI3-K family comprises 15 kinases with distinctsubstrate specificities, expression patterns, and modes of regulation(Katso et al., 2001). The class I PI3-Ks (p110α, p110β, p110δ, andp110γ) are activated by tyrosine kinases or G-protein coupled receptorsto generate PIP3, which engages downstream effectors such as theAkt/PDK1 pathway, the Tec family kinases, and the Rho family GTPases.The class II and III PI3-Ks play a key role in intracellular traffickingthrough the synthesis of PI(3)P and PI(3,4)P2. The PIKKs are proteinkinases that control cell growth (mTORC1) or monitor genomic integrity(ATM, ATR, DNA-PK, and hSmg-1).

The importance of these enzymes in diverse pathophysiology has made thePI3-K family the focus of intense interest as a new class of drugtargets (Ward et al., 2003). This interest has been fueled by the recentdiscovery that p110α is frequently mutated in primary tumors (Samuels etal., 2004) and evidence that the lipid phosphatase PTEN, an inhibitor ofPI3-K signaling, is a commonly inactivated tumor suppressor (Cantley andNeel, 1999). Efforts are underway to develop small molecule PI3-Kinhibitors for the treatment of inflammation and autoimmune disease(p110δ, p110γ, and mTOR), thrombosis (p110β), viral infection (thePIKKs) and cancer (p110α, mTOR, and others). Recently, the firstselective inhibitors of these enzymes have been reported (Camps et al.,2005; Condliffe et al., 2005; Jackson et al., 2005; Knight et al., 2004;Lau et al., 2005; Sadhu et al., 2003).

Protein tyrosine kinases, protein serine/threonine kinases, and lipidkinases are distinct classes of proteins that play critical roles inregulation and proliferation of cellular activity. Small molecules thatinhibit these protein classes have the potential to disruptdysfunctional/pathological pathways at two distinct points. For example,signaling through tyrosine kinase receptors is known to be disregulatedin several types of cancer. This signaling pathway involves downstreamproteins such as PI3 Kinase. Signaling through the serine/threonineprotein kinase mTOR (also known as the mammalian target of rapamycin) isknown to regulate cell growth, cell proliferation, cell motility, cellsurvival, protein synthesis, and transcription. Disruption of the mTORpathway is implicated as a contributing factor to various human diseaseprocesses, especially various types of cancer. An inhibitor that blocksactivity of protein tyrosine kinase and PI3 Kinase, mTOR and PI3Kinase,or mTOR, protein tyrosine kinase and PI3 Kinase, has the potential tostop the aberrant signaling at two or three different levels. Double ortriple inhibition by a small molecule may magnify drug potency,increasing the compound's therapeutic potential.

The present invention meets these and other needs in the art byproviding a new class of PI3 kinase antagonists, PI3 kinase and tryosinekinase antagonists, PI3Kinase and mTOR antagonists, and PI3Kinase, mTORand tryosine kinase antagonists.

BRIEF SUMMARY OF THE INVENTION

It has been discovered that certain compounds described herein arepotent antagonists of PI3 kinase, PI3 kinase and tryosine kinase,PI3Kinase and mTOR, or PI3Kinase, mTOR and tryosine kinase.

In one aspect, the present invention provides novel kinase antagoniststhat are PI3-Kinase affinity pocket binding antagonists (e.g. aPI3-Kinase affinity pocket binding pyrazolopyrimidine antagonist or aPI3-Kinase affinity pocket binding pyrrolopyrimidine antagonist). ThePI3-Kinase affinity pocket binding antagonist is a compound containing aPI3-Kinase affinity pocket binding moiety. The PI3-Kinase affinitypocket binding pyrazolopyrimidine antagonists of the present inventionare substituted pyrazolopyrimidine compounds containing a PI3-Kinaseaffinity pocket binding moiety. Likewise, the PI3-Kinase affinity pocketbinding pyrrolopyrimidine antagonists of the present invention aresubstituted pyrrolopyrimidine compounds containing a PI3-Kinase affinitypocket binding moiety.

In another aspect, the present invention provides the novel kinaseantagonists of Formula (I), defined below.

In another aspect, the present invention provides methods of decreasingthe catalytic activity of a PI3 Kinase (e.g. a p110δ kinase). The methodincludes the step of contacting said PI3 kinase with an activitydecreasing amount of a compound of the present invention (i.e. aPI3-Kinase affinity pocket binding antagonists, or an antagonist ofFormula I).

In another aspect, the present invention provides a method of treating acondition mediated by PI3 kinase activity, PI3 Kinase activity andtyrosine Kinase Activity, PI3 Kinase activity and mTOR activity, or PI3Kinase activity, tyrosine kinase activity, and mTOR activity in asubject in need of such treatment. The method includes administering tothe subject a therapeutically effective amount of a compound of thepresent invention (i.e. a PI3-Kinase affinity pocket bindingantagonists, or an antagonist of Formula I).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates structures of representative compounds from elevenchemotypes of PI3-K inhibitors.

FIG. 2 illustrates structures of isoform-selective PI3-K inhibitors. A.Structure of ATP in the active site of p110γ, highlighting differentregions of the ATP binding pocket. B. An alignment of all reported PI3-Kinhibitor co-crystal structures. Met 804 adopts an up conformation inall structures except PIK-39. C. Structures or models ofisoform-selective PI3-K inhibitors bound to p110γ. D. Structures ormodels of multi-targeted PI3-K inhibitors bound to p110γ.

FIG. 3 illustrates the probing of selectivity and an the PI3-Kinaseaffinity pocket. A. The structure of PIK-39 bound to p110γ suggests amodel for the binding of IC87114. PIK-293 and PIK-294 arepyrazolopyrimidine analogs of IC87114. PIK-294 projects a m-phenol intothe affinity pocket, and this compound is more potent against the classI PI3-Ks. B. (Left) Ratio of IC50 values between mutant and wild-typefor p110δ inhibitors and p110α/multi-targeted inhibitors. (Center) Doseresponse curves for binding of two p110δ inhibitors to wild-type, M7521,and M752V p110δ (Right) Models suggesting the impact of the M752I andM752V mutations in p110δ on the binding of the different classes ofinhibitors.

FIG. 4. Structures of additional PI3-K inhibitors and inactive analogs.

FIG. 5. IC50 values (μM) for selected PI3-K inhibitors against lipidkinases.

FIG. 6. Inhibition of protein kinases by PI3-K inhibitors. Valuesrepresent % activity remaining in the presence of 10 μM inhibitor.Values are average of triplicate measurements. IC50 values are inparenthesis where appropriate (μM).

FIG. 7 sets forth the sequence of a human p110δ kinase.

FIG. 8 sets forth the sequence of a human p110γ kinase.

FIG. 9 sets forth the sequence of a human p110α kinase.

FIG. 10 sets forth the sequence of a human p110β kinase.

DETAILED DESCRIPTION OF THE INVENTION I. DEFINITIONS

Abbreviations used herein have their conventional meaning within thechemical and biological arts.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents that would result from writing thestructure from right to left, e.g., —CH₂O— is equivalent to —OCH₂—.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight (i.e. unbranched) or branched chain,or cyclic hydrocarbon radical, or combination thereof, which may befully saturated, mono- or polyunsaturated and can include di- andmultivalent radicals, having the number of carbon atoms designated (i.e.C₁-C₁₀ means one to ten carbons). Examples of saturated hydrocarbonradicals include, but are not limited to, groups such as methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, forexample, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Anunsaturated alkyl group is one having one or more double bonds or triplebonds. Examples of unsaturated alkyl groups include, but are not limitedto, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkyl, as exemplified, but not limited,by —CH₂CH₂CH₂CH₂—, —CH₂CH═CHCH₂—, —CH₂C≡CCH₂—, —CH₂CH₂CH(CH₂CH₂CH₃)CH₂—.Typically, an alkyl (or alkylene) group will have from 1 to 24 carbonatoms, with those groups having 10 or fewer carbon atoms being preferredin the present invention. A “lower alkyl” or “lower alkylene” is ashorter chain alkyl or alkylene group, generally having eight or fewercarbon atoms.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcyclic hydrocarbon radical, or combinations thereof, consisting of atleast one carbon atoms and at least one heteroatom selected from thegroup consisting of O, N, P, Si and S, and wherein the nitrogen,phosphorus, and sulfur atoms may optionally be oxidized and the nitrogenheteroatom may optionally be quaternized. The heteroatom(s) O, N, P andS and Si may be placed at any interior position of the heteroalkyl groupor at the position at which alkyl group is attached to the remainder ofthe molecule. Examples include, but are not limited to, —CH₂—CH₂—O—CH₃,—CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂,—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃,—CH═CH—N(CH₃)—CH₃, O—CH₃, —O—CH₂—CH₃, and —CN. Up to two or threeheteroatoms may be consecutive, such as, for example, —CH₂—NH—OCH₃ and—CH₂—O—Si(CH₃)₃. Similarly, the term “heteroalkylene” by itself or aspart of another substituent means a divalent radical derived fromheteroalkyl, as exemplified, but not limited by, —CH₂—CH₂—S—CH₂—CH₂— and—CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylene groups, heteroatoms can alsooccupy either or both of the chain termini (e.g., alkyleneoxo,alkylenedioxo, alkyleneamino, alkylenediamino, and the like). Stillfurther, for alkylene and heteroalkylene linking groups, no orientationof the linking group is implied by the direction in which the formula ofthe linking group is written. For example, the formula —C(O)OR′—represents both —C(O)OR′— and —R′OC(O)—. As described above, heteroalkylgroups, as used herein, include those groups that are attached to theremainder of the molecule through a heteroatom, such as —C(O)R′,—C(O)NR′, —NR′R″, —OR′, —SR′, and/or —SO₂R′. Where “heteroalkyl” isrecited, followed by recitations of specific heteroalkyl groups, such as—NR′R″ or the like, it will be understood that the terms heteroalkyl and—NR′R″ are not redundant or mutually exclusive. Rather, the specificheteroalkyl groups are recited to add clarity. Thus, the term“heteroalkyl” should not be interpreted herein as excluding specificheteroalkyl groups, such as —NR′R″ or the like.

The terms “cycloalkyl” and “heterocycloalkyl”, by themselves or incombination with other terms, represent, unless otherwise stated, cyclicversions of “alkyl” and “heteroalkyl”, respectively. Additionally, forheterocycloalkyl, a heteroatom can occupy the position at which theheterocycle is attached to the remainder of the molecule. Examples ofcycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include, but are not limited to,1-1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like. The terms “cycloalkylene”and “heterocycloalkylene” refer to the divalent derivatives ofcycloalkyl and heterocycloalkyl, respectively.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” is mean to include, but not be limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, andthe like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,aromatic, hydrocarbon substituent which can be a single ring or multiplerings (preferably from 1 to 3 rings) which are fused together (e.g.naphthyl) or linked covalently. The term “heteroaryl” refers to arylgroups (or rings) that contain from one to four heteroatoms (in eachseparate ring in the case of multiple rings) selected from N, O, and S,wherein the nitrogen and sulfur atoms are optionally oxidized, and thenitrogen atom(s) are optionally quaternized. A heteroaryl group can beattached to the remainder of the molecule through a carbon orheteroatom. Non-limiting examples of aryl and heteroaryl groups includephenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl,3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl,2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl,2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl,5-indolyl, 1-isoquinolyl, 6-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl,3-quinolyl, and 6-quinolyl. Thus, the term “heteroaryl” include fusedring structures in which at least one ring includes at least two doublebonds. Substituents for each of above noted aryl and heteroaryl ringsystems are selected from the group of acceptable substituents describedbelow. The terms “arylene” and “heteroarylene” refer to the divalentradicals of aryl and heteroaryl, respectively.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxo, arylthioxo, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like) including thosealkyl groups in which a carbon atom (e.g., a methylene group) has beenreplaced by, for example, an oxygen atom (e.g., phenoxymethyl,2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like). However, theterm “haloaryl,” as used herein is meant to cover only aryls substitutedwith one or more halogens.

Where a heteroalkyl, heterocycloalkyl, or heteroaryl includes a specificnumber of members (e.g. “3 to 7 membered”), the term “member” referrersto a carbon or heteroatom.

The term “oxo” as used herein means an oxygen that is double bonded to acarbon atom.

Each of above terms (e.g., “alkyl,” “heteroalkyl,” “cycloalkyl, and“heterocycloalkyl”, “aryl,” “heteroaryl” as well as their divalentradical derivatives) are meant to include both substituted andunsubstituted forms of the indicated radical. Preferred substituents foreach type of radical are provided below.

Substituents for alkyl, heteroalkyl, cycloalkyl, heterocycloalkylmonovalent and divalent derivative radicals (including those groupsoften referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl,alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to: —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′,-halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —C(O)NR′R″,—OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)OR′,—NR—C(NR′R″)=NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —CN and—NO₂ in a number ranging from zero to (2m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″, R′″ and R″″ eachpreferably independently refer to hydrogen, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g.,aryl substituted with 1-3 halogens), substituted or unsubstituted alkyl,alkoxy or thioalkoxy groups, or arylalkyl groups. As used herein, an“alkoxy” group is an alkyl attached to the remainder of the moleculethrough a divalent oxygen radical. When a compound of the inventionincludes more than one R group, for example, each of the R groups isindependently selected as are each R′, R″, R′″ and R″″ groups when morethan one of these groups is present. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 4-, 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant toinclude, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. Fromthe above discussion of substituents, one of skill in the art willunderstand that the term “alkyl” is meant to include groups includingcarbon atoms bound to groups other than hydrogen groups, such ashaloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃,—C(O)CH₂OCH₃, and the like).

Similar to the substituents described for alkyl radicals above,exemplary substituents for aryl and heteroaryl groups (as well as theirdivalent derivatives) are varied and are selected from, for example:halogen, —OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′,—CO₂R′, —C(O)NR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″,—NR″C(O)OR′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′,—S(O)₂NR′R″, —NRSO₂R′, —CN and —NO₂, —R′, —N₃, —CH(Ph)₂,fluoro(C₁-C₄)alkoxo, and fluoro(C₁-C₄)alkyl, in a number ranging fromzero to the total number of open valences on aromatic ring system; andwhere R′, R″, R′″ and R″″ are preferably independently selected fromhydrogen, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl and substituted or unsubstituted heteroaryl. When acompound of the invention includes more than one R group, for example,each of the R groups is independently selected as are each R′, R″, R′″and R″″ groups when more than one of these groups is present.

Two of the substituents on adjacent atoms of aryl or heteroaryl ring mayoptionally form a ring of the formula -T-C(O)—(CRR′)_(q)—U—, wherein Tand U are independently —NR—, —O—, —CRR′— or a single bond, and q is aninteger of from 0 to 3. Alternatively, two of the substituents onadjacent atoms of aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula -A-(CH₂)_(r)—B—, wherein A and B areindependently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or asingle bond, and r is an integer of from 1 to 4. One of the single bondsof the new ring so formed may optionally be replaced with a double bond.Alternatively, two of the substituents on adjacent atoms of aryl orheteroaryl ring may optionally be replaced with a substituent of theformula —(CRR′)_(s)—X′—(C″R′″)_(d)—, where s and d are independentlyintegers of from 0 to 3, and X′ is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or—S(O)₂NR′—. The substituents R, R′, R″ and R′″ are preferablyindependently selected from hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl.

As used herein, the term “heteroatom” or “ring heteroatom” is meant toinclude oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), andsilicon (Si).

An “aminoalkyl” as used herein refers to an amino group covalently boundto an alkylene linker. The amino group is —NR′R″, wherein R′ and R″ aretypically selected from hydrogen, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl.

A “substituent group,” as used herein, means a group selected from thefollowing moieties:

(A) —OH, —NH₂, —SH, —CN, —CF₃, —NO₂, oxo, halogen, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and

(B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, andheteroaryl, substituted with at least one substituent selected from:

(i) oxo, —OH, —NH₂, —SH, —CN, —CF₃, —NO₂, halogen, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and

(ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, andheteroaryl, substituted with at least one substituent selected from:

(a) oxo, —OH, —NH₂, —SH, —CN, —CF₃, —NO₂, halogen, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and

(b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, orheteroaryl, substituted with at least one substituent selected from oxo,—OH, —NH₂, —SH, —CN, —CF₃, —NO₂, halogen, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, and unsubstituted heteroaryl.

A “size-limited substituent” or “size-limited substituent group,” asused herein means a group selected from all of the substituentsdescribed above for a “substituent group,” wherein each substituted orunsubstituted alkyl is a substituted or unsubstituted C₁-C₂₀ alkyl, eachsubstituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₄-C₈cycloalkyl, and each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 4 to 8 membered heterocycloalkyl.

A “lower substituent” or “lower substituent group,” as used herein meansa group selected from all of the substituents described above for a“substituent group,” wherein each substituted or unsubstituted alkyl isa substituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₅-C₇ cycloalkyl, and each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 5 to 7membered heterocycloalkyl.

The compounds of the present invention may exist as salts. The presentinvention includes such salts. Examples of applicable salt forms includehydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates,maleates, acetates, citrates, fumarates, tartrates (eg (+)-tartrates,(−)-tartrates or mixtures thereof including racemic mixtures,succinates, benzoates and salts with amino acids such as glutamic acid.These salts may be prepared by methods known to those skilled in art.Also included are base addition salts such as sodium, potassium,calcium, ammonium, organic amino, or magnesium salt, or a similar salt.When compounds of the present invention contain relatively basicfunctionalities, acid addition salts can be obtained by contacting theneutral form of such compounds with a sufficient amount of the desiredacid, either neat or in a suitable inert solvent. Examples of acceptableacid addition salts include those derived from inorganic acids likehydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived organic acids like acetic, propionic,isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric,lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like. Certain specificcompounds of the present invention contain both basic and acidicfunctionalities that allow the compounds to be converted into eitherbase or acid addition salts.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical or chiral centers) or double bonds; the enantiomers,racemates, diastereomers, tautomers, geometric isomers, stereoisometricforms that may be defined, in terms of absolute stereochemistry, as (R)—or (S)— or, as (D)- or (L)- for amino acids, and individual isomers areencompassed within the scope of the present invention. The compounds ofthe present invention do not include those which are known in art to betoo unstable to synthesize and/or isolate. The present invention ismeant to include compounds in racemic and optically pure forms.Optically active (R)— and (S)—, or (D)- and (L)-isomers may be preparedusing chiral synthons or chiral reagents, or resolved using conventionaltechniques. When the compounds described herein contain olefinic bondsor other centers of geometric asymmetry, and unless specified otherwise,it is intended that the compounds include both E and Z geometricisomers.

The term “tautomer,” as used herein, refers to one of two or morestructural isomers which exist in equilibrium and which are readilyconverted from one isomeric form to another.

It will be apparent to one skilled in the art that certain compounds ofthis invention may exist in tautomeric forms, all such tautomeric formsof the compounds being within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude all stereochemical forms of the structure; i.e., the R and Sconfigurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of atoms that constitutesuch compounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example tritium (³H), iodine-125(¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are encompassedwithin the scope of the present invention.

The term “pharmaceutically acceptable salts” is meant to include saltsof active compounds which are prepared with relatively nontoxic acids orbases, depending on the particular substituent moieties found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge et al., “Pharmaceutical Salts”, Journal ofPharmaceutical Science, 1977, 66, 1-19). Certain specific compounds ofthe present invention contain both basic and acidic functionalities thatallow the compounds to be converted into either base or acid additionsalts.

In addition to salt forms, the present invention provides compounds,which are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

The terms “a,” “an,” or “a(n)”, when used in reference to a group ofsubstituents herein, mean at least one. For example, where a compound issubstituted with “an” alkyl or aryl, the compound is optionallysubstituted with at least one alkyl and/or at least one aryl. Moreover,where a moiety is substituted with an R substituent, the group may bereferred to as “R-substituted.” Where a moiety is R-substituted, themoiety is substituted with at least one R substituent and each Rsubstituent is optionally different.

Description of compounds of the present invention are limited byprinciples of chemical bonding known to those skilled in the art.Accordingly, where a group may be substituted by one or more of a numberof substituents, such substitutions are selected so as to comply withprinciples of chemical bonding and to give compounds which are notinherently unstable and/or would be known to one of ordinary skill inthe art as likely to be unstable under ambient conditions, such asaqueous, neutral, and several known physiological conditions. Forexample, a heterocycloalkyl or heteroaryl is attached to the remainderof the molecule via a ring heteroatom in compliance with principles ofchemical bonding known to those skilled in the art thereby avoidinginherently unstable compounds.

The terms “treating” or “treatment” refers to any indicia of success inthe treatment or amelioration of an injury, pathology or condition,including any objective or subjective parameter such as abatement;remission; diminishing of symptoms or making the injury, pathology orcondition more tolerable to the patient; slowing in the rate ofdegeneration or decline; making the final point of degeneration lessdebilitating; improving a patient's physical or mental well-being. Thetreatment or amelioration of symptoms can be based on objective orsubjective parameters; including the results of a physical examination,neuropsychiatric exams, and/or a psychiatric evaluation. For example,the certain methods presented herein successfully treat cancer bydecreasing the incidence of cancer and or causing remission of cancer.

An “effective amount” is an amount sufficient to contribute to thetreatment, prevention, or reduction of a symptom or symptoms of adisease. An “effective amount” may also be referred to as a“therapeutically effective amount.” A “reduction” of a symptom orsymptoms (and grammatical equivalents of this phrase) means decreasingof the severity or frequency of the symptom(s), or elimination of thesymptom(s). A “prophylactically effective amount” of a drug is an amountof a drug that, when administered to a subject, will have the intendedprophylactic effect, e.g., preventing or delaying the onset (orreoccurrence) a disease, or reducing the likelihood of the onset (orreoccurrence) of a disease or its symptoms. The full prophylactic effectdoes not necessarily occur by administration of one dose, and may occuronly after administration of a series of doses. Thus, a prophylacticallyeffective amount may be administered in one or more administrations. An“activity decreasing amount,” as used herein, refers to an amount ofantagonist required to decrease the activity of an enzymne relateive tothe absence of the antagonist. A “function disrupting amount,” as usedherein, refers to the amount of antagonist required to disrupt thefunction of an osteoclast or leukocyte relative to the absence of theantagonist.

As used herein, the “antagonist” or “the compound of the presentinvention” refers to a compound of Formula (I), or a PI3-Kinase affinitypocket binding antagonist (e.g. a PI3-Kinase affinity pocket bindingpyrazolopyrimidine antagonists, or a PI3-Kinase affinity pocket bindingpyrrolopyrimidine antagonist). A “compound of Formula (I)” includes thecompounds of Formulae (I)-(X) as described below.

II. KINASE ANTAGONISTS

In one aspect, the present invention provides novel kinase antagonists.The kinase antagonists may be a PI3-Kinase affinity pocket bindingantagonist (e.g. a PI3-Kinase affinity pocket binding pyrazolopyrimidineantagonist, or PI3-Kinase affinity pocket binding pyrrolopyrimidineantagonist) or a compound of Formula (I). The PI3-Kinase affinity pocketbinding antagonists of the present invention are compounds containing aPI3-Kinase affinity pocket binding moiety. The PI3-Kinase affinitypocket binding pyrazolopyrimidine antagonists of the present inventionare substituted pyrazolopyrimidine compounds containing a PI3-Kinaseaffinity pocket binding moiety. Likewise, the PI3-Kinase affinity pocketbinding pyrrolopyrimidine antagonists of the present invention aresubstituted pyrrolopyrimidine compounds containing a PI3-Kinase affinitypocket binding moiety.

The PI3-Kinase affinity pocket binding moiety is a substituent which,upon contacting a p110α, p110β, p110γ, or p110δ kinase, fills spacewithin the corresponding PI3-Kinase affinity pocket. In someembodiments, the PI3-Kinase affinity pocket binding moiety displaces atleast one water molecule within the PI3-Kinase affinity pocket. ThePI3-Kinase affinity pocket binding moiety may also interact with one ormore amino acids that from part of the PI3-Kinase affinity pocket. Adescription of the PI3-Kinase affinity pocket and methods of determiningwhether a substituent fills space within the PI3-Kinase affinity pocketare set forth below.

In some embodiments, the kinase antagonist of the present invention hasthe formula:

In Formula (I), R¹ is hydrogen, halogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. R² is halogen, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl. X is═N— or ═C(H)—. R³⁶ is halogen, —NR³⁷R³⁸, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. R³⁷ and R³⁸ are independently hydrogen, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. In some embodiments, R³⁷ and R³⁸ areindependently hydrogen, or unsubstituted alkyl. R² may be a PI3-Kinaseaffinity pocket binding moiety.

In some embodiments, R³⁶ is —NH₂. Thus, the kinase antagonist may havethe formula:

In some embodiments, R¹, R², and X are as defined above in Formula (I).In certain embodiments, X is ═N—.

In some embodiments of Formulae (I) and (II), R¹ is hydrogen,R³-substituted or unsubstituted alkyl, R³-substituted or unsubstitutedheteroalkyl, R³-substituted or unsubstituted cycloalkyl, R³-substitutedor unsubstituted heterocycloalkyl, R³-substituted or unsubstituted aryl,or R³-substituted or unsubstituted heteroaryl. R² is halogen,R⁴-substituted aryl, or substituted or unsubstituted heteroaryl;

R³ is halogen, —CN, —OR⁵, —S(O)_(n)R⁶, —NR⁷R⁸, —C(O)R⁹, ═N—NH₂,—NR¹⁰—C(O)R¹¹, —NR¹²—C(O)—OR¹³, —C(O)NR¹⁴R¹⁵, —NR¹⁶S(O)₂R¹⁷, —S(O)₂NR¹⁸,R¹⁹-substituted or unsubstituted alkyl, R¹⁹-substituted or unsubstitutedheteroalkyl, R¹⁹-substituted or unsubstituted cycloalkyl,R¹⁹-substituted or unsubstituted heterocycloalkyl, R¹⁹-substituted orunsubstituted aryl, or R¹⁹-substituted or unsubstituted heteroaryl. Thensymbol n is an integer from 0 to 2.

R⁴ is halogen, —CN, —OR²⁰, —S(O)_(q)R²¹, —NR²²R²³, —C(O)R²⁴, ═N—NH₂,—NR²⁵—C(O)R²⁶, —NR²⁷—C(O)—OR²⁸, —C(O)NR²⁹R³⁰, —NR³¹S(O)₂R³², —S(O)₂NR³³,R³⁴-substituted or unsubstituted alkyl, R³⁴-substituted or unsubstitutedheteroalkyl, R³⁴-substituted or unsubstituted cycloalkyl,R³⁴-substituted or unsubstituted heterocycloalkyl, R³⁴-substituted orunsubstituted aryl, or R³⁴-substituted or unsubstituted heteroaryl. Thesymbol q represents an integer from 0 to 2.

R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R²⁰,R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², and R³³ areindependently hydrogen, R³⁵-substituted or unsubstituted alkyl,R³⁵-substituted or unsubstituted heteroalkyl, unsubstituted cycloalkyl,R³⁵-substituted or unsubstituted heterocycloalkyl, R³⁵-substituted orunsubstituted aryl, or R³⁵-substituted or unsubstituted heteroaryl. R¹⁹,R³⁴ and R³⁵ are independently hydrogen, halogen, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl.

In some embodiments, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵,R¹⁶, R¹⁷, R¹⁸, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰,R³¹, R³², and R³³ are independently hydrogen, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl. R²⁰,R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², and R³³ mayindependently be hydrogen unsubstituted alkyl, or unsubstitutedheteroalkyl.

R¹ may be R³-substituted or unsubstituted alkyl, R³-substituted orunsubstituted cycloalkyl, or R³-substituted or unsubstituted aryl. R¹may also be R³-substituted or unsubstituted alkyl, or R³-substituted orunsubstituted cycloalkyl. In some embodiments, R¹ is R³-substituted orunsubstituted C₁-C₄ alkyl, or R³-substituted or unsubstituted C₃-C₆cycloalkyl. In other embodiments, R¹ is R³-substituted or unsubstitutedC₁-C₄ alkyl, or R³-substituted or unsubstituted cyclopentyl. R¹ may alsobe methyl or unsubstituted C₃-C₆ branched alkyl (e.g. isopropyl,isobutyl, etc.).

In certain embodiments, R³ is R¹⁹-substituted or unsubstituted alkyl,R¹⁹-substituted or unsubstituted cycloalkyl, or R¹⁹-substituted orunsubstituted aryl. R³ may also be R¹⁹-substituted or unsubstitutedalkyl, R¹⁹-substituted or unsubstituted cycloalkyl, or R¹⁹-substitutedor unsubstituted aryl. In some embodiments, R³ is R¹⁹-substituted orunsubstituted alkyl, or R¹⁹-substituted or unsubstituted cycloalkyl.

R¹⁹ may be unsubstituted alkyl or unsubstituted cycloalkyl. In comeembodiments, R¹⁹ is unsubstituted C₁-C₄ alkyl or unsubstitutedcyclopentyl.

In some embodiments, R² is R⁴-substituted aryl, or R⁴-substituted orunsubstituted heteroaryl. R² may be R⁴-substituted phenyl,R⁴-substituted or unsubstituted naphthyl, R⁴-substituted orunsubstituted pyridinyl, R⁴-substituted or unsubstituted pyrimidinyl,R⁴-substituted or unsubstituted thiophenyl, R⁴-substituted orunsubstituted furanyl, R⁴-substituted or unsubstituted indolyl,R⁴-substituted or unsubstituted benzoxadiazolyl, R⁴-substituted orunsubstituted benzodioxolyl, R⁴-substituted or unsubstitutedbenzodioxanyl, R⁴-substituted or unsubstituted thianaphthanyl,R⁴-substituted or unsubstituted pyrrolopyridinyl, R⁴-substituted orunsubstituted indazolyl, R⁴-substituted or unsubstitutedtetrahydronaphthalenyl, R⁴-substituted or unsubstituted quinolinyl,R⁴-substituted or unsubstituted quinoxalinyl, R⁴-substituted orunsubstituted pyridopyrazinyl, R⁴-substituted or unsubstitutedquinazolinonyl, R⁴-substituted or unsubstituted chromenonyl,R⁴-substituted or unsubstituted benzoisoxazolyl, R⁴-substituted orunsubstituted imidazopyridinyl, R⁴-substituted or unsubstitutedbenzofuranyl, R⁴-substituted or unsubstituted dihydro-benzofuranyl,R⁴-substituted or unsubstituted dihydro-benzodioxinyl, R⁴-substituted orunsubstituted benzoimidazolonyl, or R⁴-substituted or unsubstitutedbenzothiophenyl.

In certain embodiments, R² is R⁴-substituted phenyl, R⁴-substituted orunsubstituted pyrrolepyridinyl, R⁴-substituted or unsubstitutedquinolinyl, R⁴-substituted or unsubstituted indazolyl, R⁴-substituted orunsubstituted quinolinyl indolyl, or R⁴-substituted or unsubstitutednaphthyl. R⁴ may be halogen, —CN, —OR²⁰, or —NR²²R²³. R⁴ may also simplybe halogen, or —OR²⁰.

In certain embodiments, R² is has the formula:

In Formula (III), W¹, W², W³, and W⁴ are independently ═CH—, ═CR⁴—, or═N—. Each R⁴ is as defined above in the description of Formulae (I) and(II). Ring A is a substituted or unsubstituted heteroaryl or substitutedor unsubstituted heterocycloalkyl. In some embodiments, ring A is a 6 to7 membered heterocycloalkyl or 6 to 7 membered heteroaryl. Thus, in someembodiments, ring A is partially or fully unsaturated 6- or 7-memberedring.

R²⁰ may be hydrogen or unsubstituted C₁-C₁₀ alkyl. In some embodiments,R²⁰ is hydrogen or unsubstituted C₁-C₄ alkyl. R²⁰ may also simply behydrogen or methyl.

In some embodiments, R² has the formula:

In Formulae (IV), (V) and (VI), R⁴ is absent, halogen, unsubstitutedC₁-C₄ alkyl, or —OR²⁰. The halogen may be F, Cl, or Br. In someembodiments, the halogen is F or Cl. In other embodiments, the halogenis F. R²⁰ may be hydrogen or unsubstituted C₁-C₄ alkyl.

In some embodiments, R² is 6-hydroxynaphthyl, unsubstituted 7-azaindole,unsubstituted indolyl, unsubstituted indazolyl, or unsubstitutedquinolinyl.

In some embodiments, R² has the formula:

In Formulae (VII) and (VIII), R²⁰ is as defined above. It is noted that,in accordance with the description of R²⁰ above, each R²⁰ is optionallydifferent. The symbol z is an integer from 1 to 5 (e.g. 1 or 2). In someembodiments, R²⁰ is hydrogen or unsubstituted C₁-C₁₀ alkyl (e.g. C₁-C₅alkyl such as methyl or ethyl).

In some embodiments, R² has the formula:

In Formulae (IX) and (X), above, R²⁰ is as defined above, for example,in the description of Formulae (I), (II), (VI), and (VII) above.

In some embodiments, each substituted group described above for thecompounds of the present invention is substituted with at least onesubstituent group. More specifically, in some embodiments, eachsubstituted alkyl, substituted heteroalkyl, substituted cycloalkyl,substituted heterocycloalkyl, substituted aryl, substituted heteroaryl,aryl(C₁-C₆)alkyl, and heteroaryl(C₁-C₆)alkyl described above issubstituted with at least one substituent group. In other embodiments,at least one or all of these groups are substituted with at least onesize-limited substituent group. Alternatively, at least one or all ofthese groups are substituted with at least one lower substituent group.

In other embodiments of the compounds described above, each substitutedor unsubstituted alkyl is a substituted or unsubstituted C₁-C₂₀ alkyl,each substituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₄-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 4 to 8 membered heterocycloalkyl.

Alternatively, each substituted or unsubstituted alkyl is a substitutedor unsubstituted C₁-C₈ alkyl, each substituted or unsubstitutedheteroalkyl is a substituted or unsubstituted 2 to 8 memberedheteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₅-C₇ cycloalkyl, and each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 5 to 7membered heterocycloalkyl.

In another embodiment, the compounds of Formula (I) include any or allof the compounds listed in Table 1 below.

III. THE PI3-KINASE AFFINITY POCKET

The term “PI3-Kinase affinity pocket,” as used herein, refers to acavity within p110α, p110β, p110γ, and p110δ corresponding to thelightly shaded region shown in FIGS. 2A, 2C, and 2D labeled “AffinityPocket.” FIGS. 2A, 2C, and 2D illustrate a computer model of the p110γcrystal structure. In p110γ, the surface of the PI3-Kinase affinitypocket is bound, at least in part, by the side chain of K833, D964,I879, and D841 (p110γ numbering, see FIG. 8). The surface of thecorresponding cavity in p110δ is bound, at least in part, by the sidechain of K779, D911, I825, and D787 (p110δ numbering, see FIG. 7). Thecorresponding cavity within p110α is bound, at least in part, by theside chains of K802, D933, I848, and D810 (p110α numbering, see FIG. 9).The corresponding cavity within p110β is bound, at least in part, by theside chains of K805, D937, I851, and D813 (p110β numbering, see FIG.10). The PI3-Kinase affinity pocket is not accessed by ATP.

The PI3-Kinase affinity pocket of p110δ may be referred to herein as thep110δ affinity pocket. Likewise, the PI3-Kinase affinity pocket of p110γmay be referred to herein as the p110γ affinity pocket. The PI3-Kinaseaffinity pocket includes lysine 779, which, according to computermodels, forms a hydrogen bond with the pyridine nitrogen of PIK-90 andthe phenol oxygen of PI 103 (FIG. 2D), both of which are inhibitors ofp110δ. Based on these computer modeling results, a novel antagonist wasdesigned based on the chemical structure of PIK-39 and IC87114, asdetailed below.

As shown in FIG. 2C, PIK-39 does not contain a PI3-Kinase binding pocketmoiety. And as shown in FIG. 3A, IC87114 maintains contacts to E880 andV882 in the ATP binding region of p110δ, but is also missing aPI3-Kinase binding pocket moiety. By inserting m-phenol (a PI3-Kinasebinding pocket moiety) at the C3 of the pyrazolopyrimidine of IC87114,the PI3-Kinase affinity pocket is accessed (FIG. 3A) resulting in a60-fold increase in p110δ inhibition potency.

As described above, a PI3-Kinase binding pocket moiety is a substituentwhich, upon contacting upon contacting p110α, p110β, p110γ, or p110δ,fills space within the corresponding PI3-Kinase binding pocket. Forexample, a PI3-Kinase affinity pocket binding moiety is a substituentwhich, upon contacting upon contacting p110δ, fills space within thep110α affinity pocket. Likewise, a p110α affinity pocket binding moietyis a substituent which, upon contacting upon contacting p110α, fillsspace within the p110α affinity pocket.

In some embodiments, the PI3-Kinase binding pocket moiety additionallyinteracts (e.g. bonds) with an amino acid that forms part of thePI3-Kinase binding pocket. In some related embodiments, the interactionis a hydrogen bond, van der Waals interaction, ionic bond, covalent bond(e.g. disulfide bond) or hydrophobic interaction.

IV. DETERMINING SPACE FILLING WITHIN THE PI3-KINASES AFFINITY POCKET

To determine whether the PI3-Kinase affinity pocket binding moiety fillsspace within the PI3-Kinase affinity pocket, computer modelingtechniques are employed. A query PI3-Kinase affinity pocket bindingantagonist (i.e. a test compound) is fit into a computer image of p110γ.The p110γ computer image is derived from the solved co-crystal structureof human p110γ bound to PIK-39. The PyMOL Molecular Graphics System maybe employed to generate the image. An example is presented in FIG. 3A,wherein IC87114 and PIK-294 are built into the computer image of p110γkinase, derived from the p110γ-PIK-39 co-crystal. See Knight, et al.,Cell 125: 733-745 (2006).

The computer models are typically analyzed to prevent any gross stericclashes and to satisfy key hydrogen bonds between the query PI3-Kinaseaffinity pocket binding antagonist and the p110γ protein (e.g. V882 andM804). In some embodiments, energy minimization calculations areperformed to optimize binding energy. Using these techniques, oneskilled in the art can easily determine whether a query PI3-Kinaseaffinity pocket binding antagonist includes a PI3-Kinase affinity pocketbinding moiety that fills space within the PI3-Kinase affinity pocket.

In some embodiments, the query PI3-Kinase affinity pocket bindingantagonist is analyzed to determine whether at least one bond (e.g. ahydrogen bond) is formed between the query PI3-Kinase affinity pocketbinding antagonist and an amino acid that form part of the PI3-Kinaseaffinity pocket. Using a computer modeling technique as described above,the distance between one or more amino acids that form part of thePI3-Kinase affinity pocket and a potential contact point on thePI3-Kinase affinity pocket binding moiety is determined. Based on thisdistance, one skilled in the art may determine whether at least one bondis formed between one or more amino acids that form part of thePI3-Kinase affinity pocket and a PI13-Kinase affinity pocket bindingmoiety.

V. GENERAL SYNTHESES

The compounds of the invention are synthesized by an appropriatecombination of generally well known synthetic methods. Techniques usefulin synthesizing the compounds of the invention are both readily apparentand accessible to those of skill in the relevant art. The discussionbelow is offered to illustrate certain of the diverse methods availablefor use in assembling the compounds of the invention. However, thediscussion is not intended to define the scope of reactions or reactionsequences that are useful in preparing the compounds of the presentinvention.

In Scheme I above, iodination of the pyrazolo- or pyrrolo-pyrimidine isaccomplished using an appropriate iodination reagent, such asn-iodo-succinamide. Elaboration of the 1-position may be accomplishedvia halogen displacement of a brominated substituent (e.g. a substitutedor unsubstituted alkylbromide). Palladium-catalyzed cross couplingbetween organoboronic acid and the iodo halide (i.e. Suzuki coupling),is then used to elaborate the 3-position. Recent catalyst and methoddevelopments have broadened the possible Suzuki coupling applicationsenormously, so that the scope of the reaction partners is not restrictedto aryls. Potassium trifluoroborates and organoboranes or boronateesters may be used in place of boronic acids. Some pseudohalides (forexample triflates) may also be used as coupling partners. Furtherinformation regarding Suzuki coupling may be found, for example in Kudoet al., Angew. Chem. Int. Ed. 45: 1282-1284 (2006); Kirchhoffet al., J.Am. Chem. Soc., 124: 13662-13663 (2002); Wu et al., J. Org. Chem., 68:670-673 (2003); and Molander et al., J. Org. Chem., 67: 8424-8429(2002).

VI. METHODS

In another aspect, the present invention provides methods of decreasingthe catalytic activity of a PI3 Kinase (e.g. a p110α kinase). The methodincludes the step of contacting the PI3 kinase with an activitydecreasing amount of a compound of the present invention (i.e. aPI3-Kinase affinity pocket binding antagonist or the compound of Formula(I)). In some embodiments, the antagonist is capable of decreasing thecatalytic activity of a tyrosine kinase. In some embodiments, theantagonist is a PI3-Kinase affinity pocket binding pyrazolopyrimidineantagonists, or PI3-Kinase affinity pocket binding pyrrolopyrimidineantagonists.

In some embodiments, the antagonist is specific to p110α relative to theantagonist action against p110δ, p110β, and/or p110γ. In someembodiments, the IC50 for p110α is at least 1.5, 2.0, 3.0, 4.0, 5.0, 10,20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 500, or 1000 fold lower thanthe IC50 against p110δ, p110β, and/or p110γ. In other embodiments, theIC50 of the antagonist against the p110α is less than 100 μM, 50 μM, 40μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM, 0.1 μM, 50 nM, 10 nM, 1 nM.0.5 nM, 0.1 nM, 50 pM, 10 pM, 10 pM, or 1 pM.

In some embodiments, the antagonist is specific to p110α relative to theantagonist action against insulin receptor tyrosine kinase. In someembodiments, the IC50 for p110α is at least 1.5, 2.0, 3.0, 4.0, 5.0, 10,20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 500, or 1000 fold lower thanthe IC50 against insulin receptor tyrosine kinase. In other embodiments,the IC50 of the antagonist against the p110α is less than 100 μM, 50 μM,40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM, 0.1 μM, 50 nM, 10 nM, 1nM. 0.5 nM, 0.1 nM, 50 pM, 10 pM, or 1 pM.

In some embodiments, the antagonist decreases, or is capable ofdecreasing, the catalytic activity of a tyrosine kinase. In someembodiments, the IC50 of the antagonist against the tyrosine kinase isless than 100 μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM,0.1 μM, 50 nM, 10 nM, 1 nM. 0.5 nM, 0.1 nM, 50 pM, 10 pM, or 1 pM. Sometyrosine kinases include, for example, DNA-dependent protein kinaseDNA-dependent protein kinase (pubmed protein accession number (PPAN)AAA79184), Abl tyrosine kinase (CAA52387), Bcr-Abl, hemopoietic cellkinase (PPAN CAI19695), Src (PPAN CAA24495), vascular endothelial growthfactor receptor 2 (PPAN ABB82619), vascular endothelial growth factorreceptor-2 (PPAN ABB82619), epidermal growth factor receptor (PPANAG43241), EPH receptor B4 (PPAN EAL23820), stem cell factor receptor(PPAN AAF22141), Tyrosine-protein kinase receptor TIE-2 (PPAN Q02858),fms-related tyrosine kinase 3 (PPAN NP_(—)004110), platelet-derivedgrowth factor receptor alpha (PPAN NP_(—)990080), RET (PPAN CAA73131),and functional mutants thereof. In some embodiments, the tyrosine kinaseis Abl, Bcr-Abl, EGFR, or Flt-3.

In some embodiments, the antagonist decreases, or is capable ofdecreasing, the catalytic activity of mTOR (PPAN AAI17167). In someembodiments, the IC50 of the antagonist against mTOR is less than 100μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM, 0.1 μM, 50nM, 10 nM, 1 nM. 0.5 nM, 0.1 nM, 50 pM, 10 pM, or 1 pM.

In some embodiments, the antagonist decreases, or is capable ofdecreasing, the catalytic activity of mTOR and p110α at an IC50 of lessthan 100 μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM, 0.1μM, 50 nM, 10 nM, 1 nM. 0.5 nM, 0.1 nM, 50 pM, 10 pM, or 1 pM. In otherembodiments, the antagonist decreases, or is capable of decreasing, thecatalytic activity of a tyrosine kinase and p110α at an IC50 of lessthan 100 μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM, 0.1μM, 50 nM, 10 nM, 1 nM. 0.5 nM, 0.1 nM, 50 pM, 10 pM, or 1 pM. In otherembodiments, the antagonist decreases, or is capable of decreasing, thecatalytic activity of a tyrosine kinase, mTOR, and p110α at an IC50 ofless than 100 μM, 50 μM, 40 μM, 30 μM, 20 μM, 10 μM, 5 μM, 1 μM, 0.5 μM,0.1 μM, 50 nM, 10 nM, 1 nM. 0.5 nM, 0.1 nM, 50 pM, 10 pM, or 1 pM.

In another aspect, the present invention provides a method of treating adisease or condition mediated by PI3 kinase activity, PI3 Kinaseactivity and Tyrosine Kinase Activity, PI3 Kinase activity and mTORActivity, or PI3 Kinase activity, mTOR activity, and Tyrosine KinaseActivity in a subject in need of such treatment. The method includesadministering to the subject a therapeutically effective amount of anantagonist. The antagonist is a PI3-Kinase affinity pocket bindingantagonist or the compound of Formula (I). In some embodiments theantagonist is a PI3-Kinase affinity pocket binding pyrazolopyrimidineantagonists, or PI3-Kinase affinity pocket binding pyrrolopyrimidineantagonists.

The disease may also be a bone-resorption disorder, chronic myelogenousleukemia, abnormal inflammation, autoimmune disease, thrombosis, orasthma. The disease may also be a type of cancer or cancer metastasis,including, for example, leukemia, carcinomas and sarcomas, such ascancer of the brain, breast, cervix, colon, head & neck, liver, kidney,lung, non-small cell lung, melanoma, mesothelioma, ovary, sarcoma,stomach, uterus and Medulloblastoma. Additional examples include,Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma,neuroblastoma, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis,primary macroglobulinemia, primary brain tumors, cancer, malignantpancreatic insulanoma, malignant carcinoid, urinary bladder cancer,premalignant skin lesions, testicular cancer, lymphomas, thyroid cancer,neuroblastoma, esophageal cancer, genitourinary tract cancer, malignanthypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms ofthe endocrine and exocrine pancreas, and prostate cancer. In someembodiments, the disease is selected from disease is liver cancer, coloncancer, breast cancer, melanoma, acute myelogenous leukemia, chronicmyelogenous leukemia, or nonsmall-cell lung cancer.

In another aspect, the present invention provides methods of disruptingthe function of a leukocyte or disrupting a function of an osteoclast.The method includes contacting the leukocyte or the osteoclast with afunction disrupting amount of the antagonist. The antagonist is aPI3-Kinase affinity pocket binding antagonist or the compound of Formula(I). In some embodiments the antagonist is a PI3-Kinase affinity pocketbinding pyrazolopyrimidine antagonist, or PI3-Kinase affinity pocketbinding pyrrolopyrimidine antagonist.

VII. PHARMACEUTICAL FORMULATIONS

In another aspect, the present invention provides a pharmaceuticalcomposition including an antagonist in admixture with a pharmaceuticallyacceptable excipient. One of skill in the art will recognize that thepharmaceutical compositions include the pharmaceutically acceptablesalts of the PI3-Kinase antagonists of the present invention describedabove.

In therapeutic and/or diagnostic applications, the compounds of theinvention can be formulated for a variety of modes of administration,including systemic and topical or localized administration. Techniquesand formulations generally may be found in Remington: The Science andPractice of Pharmacy (20^(th) ed.) Lippincott, Williams & Wilkins(2000).

The compounds according to the invention are effective over a widedosage range. For example, in the treatment of adult humans, dosagesfrom 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg per day, andfrom 5 to 40 mg per day are examples of dosages that may be used. A mostpreferable dosage is 10 to 30 mg per day. The exact dosage will dependupon the route of administration, the form in which the compound isadministered, the subject to be treated, the body weight of the subjectto be treated, and the preference and experience of the attendingphysician.

Pharmaceutically acceptable salts are generally well known to those ofordinary skill in the art, and may include, by way of example but notlimitation, acetate, benzenesulfonate, besylate, benzoate, bicarbonate,bitartrate, bromide, calcium edetate, carnsylate, carbonate, citrate,edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate,glutamate, glycollylarsanilate, hexykresorcinate, hydrabamine,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate,lactate, lactobionate, malate, maleate, mandelate, mesylate, mucate,napsylate, nitrate, pamoate (embonate), pantothenate,phosphate/diphosphate, polygalacturonate, salicylate, stearate,subacetate, succinate, sulfate, tannate, tartrate, or teoclate. Otherpharmaceutically acceptable salts may be found in, for example,Remington: The Science and Practice of Pharmacy (20^(th) ed.)Lippincott, Williams & Wilkins (2000). Preferred pharmaceuticallyacceptable salts include, for example, acetate, benzoate, bromide,carbonate, citrate, gluconate, hydrobromide, hydrochloride, maleate,mesylate, napsylate, pamoate (embonate), phosphate, salicylate,succinate, sulfate, or tartrate.

Depending on the specific conditions being treated, such agents may beformulated into liquid or solid dosage forms and administeredsystemically or locally. The agents may be delivered, for example, in atimed- or sustained-low release form as is known to those skilled in theart. Techniques for formulation and administration may be found inRemington: The Science and Practice of Pharmacy (20^(th) ed.)Lippincott, Williams & Wilkins (2000). Suitable routes may include oral,buccal, by inhalation spray, sublingual, rectal, transdermal, vaginal,transmucosal, nasal or intestinal administration; parenteral delivery,including intramuscular, subcutaneous, intramedullary injections, aswell as intrathecal, direct intraventricular, intravenous,intra-articullar, intra-sternal, intra-synovial, intra-hepatic,intralesional, intracranial, intraperitoneal, intranasal, or intraocularinjections or other modes of delivery.

For injection, the agents of the invention may be formulated and dilutedin aqueous solutions, such as in physiologically compatible buffers suchas Hank's solution, Ringer's solution, or physiological saline buffer.For such transmucosal administration, penetrants appropriate to thebarrier to be permeated are used in the formulation. Such penetrants aregenerally known in the art.

Use of pharmaceutically acceptable inert carriers to formulate thecompounds herein disclosed for the practice of the invention intodosages suitable for systemic administration is within the scope of theinvention. With proper choice of carrier and suitable manufacturingpractice, the compositions of the present invention, in particular,those formulated as solutions, may be administered parenterally, such asby intravenous injection. The compounds can be formulated readily usingpharmaceutically acceptable carriers well known in the art into dosagessuitable for oral administration. Such carriers enable the compounds ofthe invention to be formulated as tablets, pills, capsules, liquids,gels, syrups, slurries, suspensions and the like, for oral ingestion bya subject (e.g. patient) to be treated.

For nasal or inhalation delivery, the agents of the invention may alsobe formulated by methods known to those of skill in the art, and mayinclude, for example, but not limited to, examples of solubilizing,diluting, or dispersing substances such as, saline, preservatives, suchas benzyl alcohol, absorption promoters, and fluorocarbons.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in aneffective amount to achieve its intended purpose. Determination of theeffective amounts is well within the capability of those skilled in theart, especially in light of the detailed disclosure provided herein.

In addition to the active ingredients, these pharmaceutical compositionsmay contain suitable pharmaceutically acceptable carriers comprisingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Thepreparations formulated for oral administration may be in the form oftablets, dragees, capsules, or solutions.

Pharmaceutical preparations for oral use can be obtained by combiningthe active compounds with solid excipients, optionally grinding aresulting mixture, and processing the mixture of granules, after addingsuitable auxiliaries, if desired, to obtain tablets or dragee cores.Suitable excipients are, in particular, fillers such as sugars,including lactose, sucrose, mannitol, or sorbitol; cellulosepreparations, for example, maize starch, wheat starch, rice starch,potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethyl-cellulose (CMC),and/or polyvinylpyrrolidone (PVP: povidone). If desired, disintegratingagents may be added, such-as the cross-linked polyvinylpyrrolidone,agar, or alginic acid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethyleneglycol (PEG), and/or titanium dioxide, lacquer solutions, and suitableorganic solvents or solvent mixtures. Dye-stuffs or pigments may beadded to the tablets or dragee coatings for identification or tocharacterize different combinations of active compound doses.

Pharmaceutical preparations that can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin, and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols (PEGs). In addition, stabilizers may be added.

Depending upon the particular condition, or disease state, to be treatedor prevented, additional therapeutic agents, which are normallyadministered to treat or prevent that condition, may be administeredtogether with the inhibitors of this invention. For example,chemotherapeutic agents or other anti-proliferative agents may becombined with the inhibitors of this invention to treat proliferativediseases and cancer. Examples of known chemotherapeutic agents include,but are not limited to, adriamycin, dexamethasone, vincristine,cyclophosphamide, fluorouracil, topotecan, taxol, interferons, andplatinum derivatives.

Other examples of a gents the inhibitors of this invention may also becombined with include, without limitation, anti-inflammatory agents suchas corticosteroids, TNF blockers, IL-1 RA, azathioprine,cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; agents for treating diabetessuch as insulin, insulin analogues, alpha glucosidase inhibitors,biguanides, and insulin sensitizers; and agents for treatingimmunodeficiency disorders such as gamma globulin.

These additional agents may be administered separately, as part of amultiple dosage regimen, from the composition. Alternatively, theseagents may be part of a single dosage form, mixed together with theinhibitor in a single composition.

The present invention is not to be limited in scope by the exemplifiedembodiments, which are intended as illustrations of single aspects ofthe invention. Indeed, various modifications of the invention inaddition to those described herein will become apparent to those havingskill in the art from the foregoing description. Such modifications areintended to fall within the scope of the invention. Moreover, any one ormore features of any embodiment of the invention may be combined withany one or more other features of any other embodiment of the invention,without departing from the scope of the invention. For example, thePI3-Kinase antagonists of the present invention described above areequally applicable to the methods of treatment and methods of inhibitingkinases described herein. References cited throughout this applicationare examples of the level of skill in the art and are herebyincorporated by reference herein in their entirety for all purposes,whether previously specifically incorporated or not.

VIII. EXAMPLES

The following examples are meant to illustrate certain embodiments ofthe invention, and not to limit the scope of the invention.

General Methods. All chemicals, reagents and solvents used werepurchased commercially and used as received. dH2O refers to deioinizedwater. Evaporation of solvents was carried out on a rotary evaporatorunder reduced pressure. Compounds were purified by High Pressure LiquidChromatography (HPLC) eluting with dH₂O-MeCN-trifluroacetic acid,50:50:0.1, unless otherwise indicated. Analysis of products was carriedout on a Liquid Chromatography Mass Spectrometer (LCMS) using MeCN-0.1%formic acid (varying ratios) as eluent.

A. Selected Reaction Procedures.

Synthesis of 1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA18). A solution of250 mL of formamide and 3-amino-4-pyrazolecarbonitrile (25 g, 0.231 mol)was heated to 180° C. overnight under an argon atmosphere. Reaction wascooled and 400 mL of dH₂O were added. The resulting solid was filteredand rinsed with cold dH₂O. White solid precipitate was collected anddried in vacuo overnight to yield BA18 (39 g, 100% yield). ESI-MS (M+H)⁺m/z calcd 136.1, found 136.1.

Synthesis of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA19). Asolution of 3H-pyrazolo[3,4-d]pyrimidin-4-amine (10 g, 0.074 mol) andn-iodo-succinamide (25 g, 0.111 mol) in DMF (80 mL) was heated to 80° C.overnight under an argon atmosphere. The resulting solid was filteredand rinsed with cold EtOH. Product was dried in vacuo overnight to yieldBA19 (24 g, 100% yield). ESI-MS (M+H)⁺ m/z calcd 262.0, found 262.0

Synthesis of 3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA12). A solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2 g,0.0077 mol) and K₂CO₃ (4.2 g, 0.031 mol) in DMF (50 mL) was brought to80° C. under an argon atmosphere. Isopropylbromide (1.0 g, 0.0084 mol)was added with a syringe. Reaction was refluxed under argon atmospherefor 2 hours. Solid K₂CO₃ was removed by filtration. Solvent waspartially removed in vacuo. Sodium citrate (50 mL) was added andreaction was extracted with EtOAc. Organic phases concentrated in vacuoand purified using silica gel column chromatography [MeOH—H₂Cl₂, 5:95]yielding BA12 (1.68 g, 72% yield). ESI-MS (M+H)⁺ m/z calcd 304.0, found304.1.

General Suzuki coupling. Preparation of final products (see Table 1 forfinal product names and structures).

3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol, 1 equivalent) was dissolved in DME (12 ml). Boronic acid (1.1equivalent) was dissolved in EtOH (3.3 ml) and added to reactionmixture. Pd(PPh3)4 (30 mg, 0.026 mmol, 0.2 equivalents) and saturatedNa2CO3 (1.9 ml) were added to the reaction mixture and heated to 80° C.under argon and refluxed for 8 hours. After cooling, the reaction wasextracted with saturated NaCl and CH2Cl2. Organic phases were combinedand solvent was removed. Resulting solid (or oil) was dissolved indH2O-MeCN-trifluroacetic acid, 50:50:0.1 and purified by HPLC. Purifiedproduct (varying yields) was confirmed by LCMS.

Synthesis of4-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-benzenesulfonamide(BA14). A solution of benzenesulfonamide-4-boronic acid pinacol ester(23 mg, 0.08 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA14 (2.2 mg,10% yield). ESI-MS (M+H)⁺ m/z calcd 333.1, found 333.1.

Synthesis of1-isopropyl-3-(3-methoxy-4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA15). A solution of 2methoxy-4-(4,4,5,5-tetramethyl-1,3-2-dioxaborolan-2-yl) phenol (19 mg,0.08 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA15 (4.3 mg,20% yield). ESI-MS (M+H)⁺ m/z calcd 300.1, found 300.2.

Synthesis of6-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)naphthalen-2-ol(BA17). A solution of 6-hydroxynaphthalen-2-yl-2-boronic acid (15 mg,0.08 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA15 (4.8 mg,23% yield). ESI-MS (M+H)⁺ m/z calcd 320.1, found 320.1.

Synthesis of tert-butyl4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenylcarbamate(BA20). A solution of 4 4-N-Boc-amino-3-methoxy-benzeneboronic acid (48mg, 0.18 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg, 0.18mmol) in DME (12 mL). Pd(PPh₃)₄ (40 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA20. ESI-MS(M+H)⁺ m/z calcd 399.2, found 399.1.

Synthesis of3-(4-amino-3-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA20d). A solution of tert-butyl4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenylcarbamate(BA20) (20 mg, 0.05 mmol) in CH₂Cl₂, TFA, S(CH₂)₂, H₂O (45:45:5:5) (1mL) was stirred at room temperature for 15 minutes. NaHCO₃ (2 mL) wasadded till reaction was alkaline. Reaction was extracted with H₂O andCH₂Cl₂. Organic phases were combined, concentrated in vacuo and purifiedby RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA20d.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)pyridine-2-carbonitrile(BA21). A solution of 2-cyanopyridine 5-boronic acid pinocol ester (18mg, 0.08 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA21 (2.5 mg,14% yield). ESI-MS (M+H)⁺ m/z calcd 280.1, found 280.1.

Synthesis of3-(3-(benzyloxy)-5-fluorophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine.A solution of (3-Benzyloxy-5-fluorophenyl)boronic acid (29 mg, 5.80mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA22 (15.6mg, 60% yield). ESI-MS (M+H)⁺ m/z calcd 378.1, found 378.0.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5-fluorophenol(BA22). A solution of-(3-(benzyloxy)-5-fluorophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(15 mg, 0.04 mmol) in MeOH (0.9 mL) was flushed with argon. Pd onactivated carbon (10 mL) was carefully added while keeping reactionunder an argon atmosphere. Reaction was flushed with H₂ gas and leftunder H₂ atmosphere overnight at room temperature. The reaction wasfiltered through celite and rinsed with MeOH to yield BA22 (15 mg, 100%yield). ESI-MS (M+H)⁺ m/z calcd 288.1, found 288.1.

Synthesis of1-isopropyl-3-(3,4-dimethoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA23). A solution of 3,4-Dimethoxyphenylboronic acid (24 mg, 0.13 mmol)in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA23 (13.1mg, 60% yield). ESI-MS (M+H)⁺ m/z calcd 314.0, found 314.1.

Synthesis of(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)methanol(BA26). A solution of (3-Hydroxymethylphenyl)boronic acid (24 mg, 0.13mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA26 (8.4 mg,42% yield). ESI-MS (M+H)⁺ m/z calcd 283.1, found 284.2.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4,5-dihydrothiazol-2-yl)benzamide(BA30). A solution of[3-((4,5-dihydrothiazol-2-yl)carbamoyl)phenyl]boronic acid (19 mg, 0.08mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA30 (17.8mg, 67% yield).

Synthesis of1-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)ethanone(BA31). A solution of 4-Acetylphenylboronic acid (12.7 mg, 0.08 mmol) inEtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA31 (12.9mg, 62% yield).

Synthesis of(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)methanol(BA32). A solution of (4-Aminocarbonyl-3-chlorophenyl)boronic acid (16mg, 0.08 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA32 (9.7 mg,42% yield). ESI-MS (M+H)⁺ m/z calcd 331.1, found 331.1.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-3-methylthiophene-2-carbaldehyde(BA34). A solution of 5-Formyl-3-methylthiophene-2-boronic acid (26 mg,0.14 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA34 (14.7mg, 38% yield). ESI-MS (M+H)⁺ m/z calcd 302.1, found 302.0.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)furan-3-carbaldehyde(BA35). A solution of 4-Formylfuran-2-boronic acid (20 mg, 0.14 mmol) inEtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA35 (13.5mg, 39% yield). ESI-MS (M+H)⁺ m/z calcd 272.1, found 272.1.

Synthesis ofN-[3-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-phenyl]-methanesulfonamide(BA38). A solution of 3-Methanesulfonylaminophenylboronic acid (32 mg,0.15 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (16 mg, 0.014 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA38 (24.3mg, 54% yield). ESI-MS (M+H)⁺ m/z calcd 347.1, found 347.0.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzonitrile(BA39). A solution of 3-Cyanophenylboronic acid (23 mg, 0.15 mmol) inEtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA39 (14.9mg, 41% yield). ESI-MS (M+H)⁺ m/z calcd 279.1, found 279.0.

Synthesis ofN-[4-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-phenyl]-methanesulfonamide(BA40). A solution of 4-Methanesulfonylaminophenylboronic acid (24 mg,0.11 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA40 (0.9 mg,3% yield). ESI-MS (M+H)⁺ m/z calcd 347.1, found 347.0.

Synthesis of3-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-benzenesulfonamide(BA41). A solution of Benzenesulfonamide-3-boronic acid pinacol ester(31 mg, 0.11 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA41 (9.2 mg,28% yield). ESI-MS (M+H)⁺ m/z calcd 333.1, found 333.0.

Synthesis of2-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzo[b]thiophene-5-carbaldehyde(BA42). A solution of 5-Formylbenzo[b]thiophene-2-boronic acid pinacolester (31 mg, 0.11 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA42 (15.2mg, 45% yield). ESI-MS (M+H)⁺ m/z calcd 338.1, found 338.0.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-3-carbaldehyde(BA43). A solution of N-Boc-3-formyl-5-indoleboronic acid pinacol ester(40 mg, 0.11 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA). The TFA frompurification hydrolyzed the Boc to yield BA43. ESI-MS (M+H)⁺ m/z calcd321.1, found 321.0.

Synthesis of3-(benzo[c][1,2,5]oxadiazol-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA44). A solution of Benzo[c][1,2,5]oxadiazole-5-boronic acid (18 mg,0.11 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA44. ESI-MS(M+H)⁺ m/z calcd 296.1, found 296.1.

Synthesis of2-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)acetonitrile(BA45). A solution of (4-Cyanomethylphenyl)boronic acid (18 mg, 0.11mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA45. ESI-MS(M+H)⁺ m/z calcd 293.1, found 293.1.

Synthesis of2-(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)acetonitrile(BA46). A solution of (3-Cyanomethylphenyl)boronic acid (18 mg, 0.11mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA45. ESI-MS(M+H)⁺ m/z calcd 293.1, found 293.1.

Synthesis of1-isopropyl-3-(4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA48). A solution of (4-Methoxyphenylboronic acid (17 mg, 0.11 mmol) inEtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA48 (4.5 mg,16% yield). ESI-MS (M+H)⁺ m/z calcd 284.1, found 284.1.

Synthesis of1-isopropyl-3-(3-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA49). A solution of 3-Methoxyphenylboronic acid (17 mg, 0.11 mmol) inEtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA49. ESI-MS(M+H)⁺ m/z calcd 284.1, found 284.0.

Synthesis of1-isopropyl-3-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA52).A solution of 3-Pyridinylboronic acid (15 mg, 0.14 mmol) in EtOH (3.3mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (15 mg, 0.015 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O) to yield BA52. ESI-MS (M+H)⁺m/z calcd 255.1, found 255.0.

Synthesis of1-isopropyl-3-(pyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA53). A solution of 5-Pyrimidinylboronic acid (15 mg, 0.14 mmol) inEtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (15 mg, 0.015 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O) to yield BA53. ESI-MS (M+H)⁺m/z calcd 256.1, found 256.1.

Synthesis of3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA54). A solution of 2,3-dihydro-1,4-benzodioxin-6-ylboronic acid (26mg, 0.14 mmol) in EtOH (3.3 mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA54 (6 mg,15% yield). ESI-MS (M+H)⁺ m/z calcd 312.1, found 312.0.

Synthesis of1-(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)ethanone(BA55). A solution of 3-Acetylphenylboronic acid (23 mg, 0.14 mmol) inEtOH (3.3-mL) was-added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA55 (7 mg,18% yield). ESI-MS (M+H)⁺ m/z calcd 296.1, found 296.1.

Synthesis of4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol (BA56). Asolution of 4-Hydroxyphenylboronic acid (30 mg, 0.14 mmol) in EtOH (3.3mL) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 mL). Pd(PPh₃)₄ (30 mg, 0.03 mmol) and saturated Na₂CO₃(1.9 mL) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH₂Cl₂. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield BA56 (12 mg,32% yield). ESI-MS (M+H)⁺ m/z calcd 270.1, found 270.1.

Synthesis of PI3-K/Tyrosine Kinase Dual Inhibitors

Synthesis of 1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA18). A solution of250 ml of formamide and 3-amino-4-pyrazolecarbonitrile (25 g, 0.231 mol)was heated to 180° C. overnight under an argon atmosphere. Reaction wascooled and 400 ml of dH2O were added. The resulting solid was filteredand rinsed with cold dH2O. White solid precipitate was collected anddried in vacuo overnight to yield BA18 (39 g, 100% yield). ESI-MS (M+H)+m/z calcd 136.1, found 136.1.

Synthesis of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA19). Asolution of 3H-pyrazolo[3,4-d]pyrimidin-4-amine (10 g, 0.074 mol) andn-iodo-succinamide (25 g, 0.111 mol) in DMF (80 ml) was heated to 80° C.overnight under an argon atmosphere. The resulting solid was filteredand rinsed with cold EtOH. Product was dried in vacuo overnight to yieldBA19 (24 g, 100% yield). ESI-MS (M+H)+ m/z calcd 262.0, found 262.0

Synthesis of 3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA12). A solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2 g,0.0077 mol) and K2CO3 (4.2g, 0.031 mol) in DMF (50 ml) was brought to80° C. under an argon atomosphere. Isopropylbromide (1.0 g, 0.0084 mol)was added with a syringe. Reaction was refluxed under argon atmospherefor 2 hours. Solid K2CO3 was removed by filtration. Solvent waspartially removed in vacuo. Sodium citrate (50 ml) was added andreaction was extracted with EtOAc. Organic phases concentrated in vacuoand purified using silica gel column chromatography [MeOH—CH2Cl2, 5:95]yielding BA12 (1.68 g, 72% yield). ESI-MS (M+H)+ m/z calcd 304.0, found304.1.

Synthesis of4-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-benzenesulfonamide(BA14). A solution of benzenesulfonamide-4-boronic acid pinacol ester(23 mg, 0.08 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA14 (2.2 mg,10% yield). ESI-MS (M+H)+ m/z calcd 333.1, found 333.1.

Synthesis of1-isopropyl-3-(3-methoxy-4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA15). A solution of 2methoxy-4-(4,4,5,5-tetramethyl-1,3-2-dioxaborolan-2-yl) phenol (19 mg,0.08 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA15 (4.3 mg,20% yield). ESI-MS (M+H)+ m/z calcd 300.1, found 300.2.

Synthesis of6-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)naphthalen-2-ol(BA17). A solution of 6-hydroxynaphthalen-2-yl-2-boronic acid (15 mg,0.08 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA15 (4.8 mg,23% yield). ESI-MS (M+H)+ m/z calcd 320.1, found 320.1.

Synthesis of tert-butyl4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenylcarbamate(BA20). A solution of 4 4-N-Boc-amino-3-methoxy-benzeneboronic acid (48mg, 0.18 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg, 0.18mmol) in DME (12 ml). Pd(PPh3)4 (40 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA20. ESI-MS(M+H)+ m/z calcd 399.2, found 399.1.

Synthesis of3-(4-amino-3-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA20d). A solution of tert-butyl4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenylcarbamate(BA20) (20 mg, 0.05 mmol) in CH2Cl2, TFA, S(CH2)2, H2O (45:45:5:5) (1ml) was stirred at room temperature for 15 minutes. NaHCO3 (2 ml) wasadded till reaction was alkaline. Reaction was extrated with H2O andCH2Cl2. Organic phases were combined, concentrated in vacuo and purifiedby RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA20d.

Synthesis of2-amino-5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol(BA20dd). BA20 (tert-butyl4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenylcarbamate,7 mg, 0.018 mmol) was dissolved in CH2Cl2 (2.5 ml) and stirred under anargon atmosphere at room temperature. BBr3 (0.500 ml) was added slowlywith a syringe. The reaction mixture was stirred overnight, under argonat room temperature. BBr3 was removed in vacuo and the remaining solidwas purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA20dd.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)pyridine-2-carbonitrile(BA21). A solution of 2-cyanopyridine 5-boronic acid pinocol ester (18mg, 0.08 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA21 (2.5 mg,14% yield). ESI-MS (M+H)+ m/z calcd 280.1, found 280.1.

Synthesis of3-(3-(benzyloxy)-5-fluorophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine.A solution of (3-Benzyloxy-5-fluorophenyl)boronic acid (29 mg, 5.80mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA22 (15.6mg, 60% yield). ESI-MS (M+H)+ m/z calcd 378.1, found 378.0.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5-fluorophenol(BA22). A solution of-(3-(benzyloxy)-5-fluorophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(15 mg, 0.04 mmol) in MeOH (0.9 ml) was flushed with argon. Pd onactivated carbon (10 ml) was carefully added while keeping reactionunder an argon atmosphere. Reaction was flushed with H2 gas and leftunder H2 atmosphere overnight at room temperature. The reaction wasfiltered through celite and rinsed with MeOH to yield BA22 (15 mg, 100%yield). ESI-MS (M+H)+ m/z calcd 288.1, found 288.1.

Synthesis of1-isopropyl-3-(3,4-dimethoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA23). A solution of 3,4-Dimethoxyphenylboronic acid (24 mg, 0.13 mmol)in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA23 (13.1mg, 60% yield). ESI-MS (M+H)+ m/z calcd 314.0, found 314.1.

Synthesis of tert-butyl2-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5-(benzyloxy)-1H-indole-1-carboxylate(BA24). A solution of 5-Benzyloxy-1-BOC-indole-2-boronic acid (303 mg,0.83 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (100 mg, 0.33mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [EtOAc-hexanes,5:95] to yield BA24. ESI-MS (M+H)+ m/z calcd 499.2, found 499.2.

Synthesis of2-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-5-ol(BA24dd). BA24(3-(4-fluoro-3-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine,30 mg, 0.10 mmol) was dissolved in a solution of formic acid (4.5 ml, 10equivalents) and HCl (0.45 ml, 1 equivalent). The reaction was heatedand stirred for one hour under an argon atmosphere. The reaction wasthen concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA)to yield BA24dd. ESI-MS (M+H)+ m/z calcd 309.1, found 309.1.

Synthesis of(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)methanol(BA26). A solution of (3-Hydroxymethylphenyl)boronic acid (24 mg, 0.13mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA26 (8.4 mg,42% yield). ESI-MS (M+H)+ m/z calcd 283.1, found 284.2.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4,5-dihydrothiazol-2-yl)benzamide(BA30). A solution of[3-((4,5-dihydrothiazol-2-yl)carbamoyl)phenyl]boronic acid (19 mg, 0.08mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA30 (17.8mg, 67% yield).

Synthesis of1-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)ethanone(BA31). A solution of 4-Acetylphenylboronic acid (12.7 mg, 0.08 mmol) inEtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA31 (12.9mg, 62% yield).

Synthesis of(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)methanol(BA32). A solution of (4-Aminocarbonyl-3-chlorophenyl)boronic acid (16mg, 0.08 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20 mg, 0.07mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA32 (9.7 mg,42% yield). ESI-MS (M+H)+ m/z calcd 331.1, found 331.1.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-3-methylthiophene-2-carbaldehyde(BA34). A solution of 5-Formyl-3-methylthiophene-2-boronic acid (26 mg,0.14 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA34 (14.7mg, 38% yield). ESI-MS (M+H)+ m/z calcd 302.1, found 302.0.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)furan-3-carbaldehyde(BA35). A solution of 4-Formylfuran-2-boronic acid (20 mg, 0.14 mmol) inEtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA35 (13.5mg, 39% yield). ESI-MS (M+H)+ m/z calcd 272.1, found 272.1.

Synthesis ofN-[3-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-phenyl]-methanesulfonamide(BA38). A solution of 3-Methanesulfonylaminophenylboronic acid (32 mg,0.15 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (16 mg, 0.014 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA38 (24.3mg, 54% yield). ESI-MS (M+H)+ m/z calcd 347.1, found 347.0.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzonitrile(BA39). A solution of 3-Cyanophenylboronic acid (23 mg, 0.15 mmol) inEtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA39 (14.9mg, 41% yield). ESI-MS (M+H)+ m/z calcd 279.1, found 279.0.

Synthesis ofN-[4-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-phenyl]-methanesulfonamide(BA40). A solution of 4-Methanesulfonylaminophenylboronic acid (24 mg,0.11 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA40 (0.9 mg,3% yield). ESI-MS (M+H)+ m/z calcd 347.1, found 347.0.

Synthesis of3-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-benzenesulfonamide(BA41). A solution of Benzenesulfonamide-3-boronic acid pinacol ester(31 mg, 0.11 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA41 (9.2 mg,28% yield). ESI-MS (M+H)+ m/z calcd 333.1, found 333.0.

Synthesis of2-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzo[b]thiophene-5-carbaldehyde(BA42). A solution of 5-Formylbenzo[b]thiophene-2-boronic acid pinacolester (31 mg, 0.11 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA42 (15.2mg, 45% yield). ESI-MS (M+H)+ m/z calcd 338.1, found 338.0.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-3-carbaldehyde(BA43). A solution of N-Boc-3-formyl-5-indoleboronic acid pinacol ester(40 mg, 0.11 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA). The TFA frompurification hydrolyzed the Boc to yield BA43. ESI-MS (M+H)+ m/z calcd321.1, found 321.0.

Synthesis of3-(benzo[c][1,2,5]oxadiazol-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA44). A solution of Benzo[c][1,2,5]oxadiazole-5-boronic acid (18 mg,0.11 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA44. ESI-MS(M+H)+ m/z calcd 296.1, found 296.1.

Synthesis of2-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)acetonitrile(BA45). A solution of (4-Cyanomethylphenyl)boronic acid (18 mg, 0.11mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H20:0.1% TFA) to yield BA45. ESI-MS(M+H)+ m/z calcd 293.1, found 293.1.

Synthesis of2-(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)acetonitrile(BA46). A solution of (3-Cyanomethylphenyl)boronic acid (18 mg, 0.11mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA45. ESI-MS(M+H)+ m/z calcd 293.1, found 293.1.

Synthesis of1-isopropyl-3-(4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA48). A solution of (4-Methoxyphenylboronic acid (17 mg, 0.11 mmol) inEtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA48 (4.5 mg,16% yield). ESI-MS (M+H)+ m/z calcd 284.1, found 284.1.

Synthesis of1-isopropyl-3-(3-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA49). A solution of 3-Methoxyphenylboronic acid (17 mg, 0.11 mmol) inEtOH (3.3 ml) was added to a solution-of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.10mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight: After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA49. ESI-MS(M+H)+ m/z calcd 284.1, found 284.0.

Synthesis of1-isopropyl-3-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA52).A solution of 3-Pyridinylboronic acid (15 mg, 0.14 mmol) in EtOH (3.3ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (15 mg, 0.015 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O) to yield BA52. ESI-MS (M+H)+m/z calcd 255.1, found 255.0.

Synthesis of1-isopropyl-3-(pyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA53). A solution of 5-Pyrimidinylboronic acid (15 mg, 0.14 mmol) inEtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (15 mg, 0.015 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O) to yield BA53. ESI-MS (M+H)+m/z calcd 256.1, found 256.1.

Synthesis of3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA54). A solution of 2,3-dihydro-1,4-benzodioxin-6-ylboronic acid (26mg, 0.14 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA54 (6 mg,15% yield). ESI-MS (M+H)+ m/z calcd 312.1, found 312.0.

Synthesis of1-(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)ethanone(BA55). A solution of 3-Acetylphenylboronic acid (23 mg, 0.14 mmol) inEtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA55 (7 mg,18% yield). ESI-MS (M+H)+ m/z calcd 296.1, found 296.1.

Synthesis of4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol (BA56). Asolution of 4-Hydroxyphenylboronic acid (30 mg, 0.14 mmol) in EtOH (3.3ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (40 mg, 0.13mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA56 (12 mg,32% yield). ESI-MS (M+H)+ m/z calcd 270.1, found 270.1.

Synthesis of4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenol(BA59). A solution of 3-fluoro-4-hydroxyphenylboronic acid (103 mg, 0.66mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (100 mg, 0.33mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP using silica gel column chromatography[MeOH—CH2Cl2, 2:98] to yield BA59 (26 mg, 27% yield). ESI-MS (M+H)+ m/zcalcd 288, found 288.

Synthesis of4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-3-methylphenol(BA60). A solution of 4-hydroxy-2-methylphenylboronic acid (110 mg, 0.66mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (100 mg, 0.33mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [MeOH—CH2Cl2,2:98] to yield BA60 (42 mg, 22% yield). ESI-MS (M+H)+ m/z calcd 284,found 284.

Synthesis of3-(4-fluoro-3-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA62). A solution of 4-fluoro-3-methoxyphenylboronic acid (61 mg, 0.36mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (90 mg, 0.30mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [MeOH—CH2Cl2,2:98] to yield BA62 (40 mg, 44% yield). ESI-MS (M+H)+ m/z calcd 302,found 302.

Synthesis of5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenol(BA62d). A solution of BA62(3-(4-fluoro-3-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine,30 mg, 0.10 mmol) was dissolved in CH2Cl2 (5 ml) and stirred under anargon atmosphere. BBr3 (500 uL, 0.5 mol) was added slowly with asyringe, while stirring. The reaction was stirred at room temperaturefor 3 hours then concentrated in vacuo and purified using silica gelcolumn chromatography [MeOH—CH2Cl2, 2:98] to yield BA62d (23 mg, 44%yield). ESI-MS (M+H)+ m/z calcd 288.1, found 288.1.

Synthesis of3-(2,5-difluoro-4-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA63). A solution of 2,5-difluoro-4-methoxyphenylboronic acid (84 mg,0.45 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (54 mg, 0.18mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [MeOH—CH2Cl2,2:98] to yield BA63 (50 mg, 17% yield). ESI-MS (M+H)+ m/z calcd 320.1,found 320.0.

Synthesis of4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2,5-difluorophenol(BA93).3-(2,5-difluoro-4-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA63) (20 mg, 0.06 mmol) was dissolved in CH2Cl2 (2 ml) and BBr3 (0.630mL, 0.63 mmol) was added slowly with a syringe, while stirring. Thereaction was stirred at room temperature for overnight then concentratedin vacuo and purified using by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA93(6.7 mg, 35% yield). ESI-MS (M+H)+ m/z calcd 306.1, found 306.0.

Synthesis of1-isopropyl-3-(3,4,5-trimethoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA64). A solution of 3,4,5-trimethoxyphenylboronic acid (123 mg, 0.58mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (70 mg, 0.23mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [MeOH—CH2Cl2,2:98] to yield BA64 (70 mg, 89% yield). ESI-MS (M+H)+ m/z calcd 344.1,found 344.0.

Synthesis of1-isopropyl-3-(2,3-dimethoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA65). A solution 2,3-dimethoxyphenylboronic acid (105 mg, 0.58 mmol)in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (70 mg, 0.23mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [MeOH—CH2Cl2,2:98] to yield BA65 (63 mg, 88% yield). ESI-MS (M+H)+ m/z calcd 314.1,found 314.1.

Synthesis of1-isopropyl-3-(2,4-dimethoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA66). A solution 2,4-dimethoxypyrimidin-5-yl-5-boronic acid (106 mg,0.58 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (70 mg, 0.23mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by silica gel column chromatography [MeOH—CH2Cl2,2:98] to yield BA66. ESI-MS (M+H)+ m/z calcd 316.1, found 316.0.

Synthesis of1-cyclopentyl-3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA79). 3-fluoro-5-methoxybenzoic acid (5 g, 0.029 mol) was stirred inCH2Cl2 (50ml) at 0° C. under an argon atmosphere. DMF (9 drops,catalytic) was added, followed by oxalyl chloride (12.7 ml, 0.147 mol).Reaction was warmed to room temperature then stirred under argon for onehour. Reaction was concentrated in vacuo to yield3-fluoro-5-methoxybenzoyl chloride (BA67).

A solution of malononitrile (2.87 g, 0.044 mmol) in dry THF (50 ml) wasstirred at 0° C. under an argon atmosphere. NaH in paraffin oil (4.64 g,0.116 mol) was added piece-wise to solution. 3-fluoro-5-methoxybenzoylchloride (BA67, 0.029 mol) was dissolved in 50 ml dry THF and addedslowly to reaction. Reaction was warmed to room temperature and stirredunder argon for 24 hours. 1N HCl (200 ml) was slowly added, thenreaction mixture was extracted with EtOAc. Organic phases were combined,dried with magnesium sulfate, then concentrated in vacuo to yield2-((3-fluoro-5-methoxyphenyl)(methoxy)methylene)malononitrile (BA69).

2-((3-fluoro-5-methoxyphenyl)(methoxy)methylene)malononitrile (BA69,0.029 mol) stirred in EtOH (20 ml) at room temperature under an argonatmosphere. Hydrazine (1.4 ml, 29 mmol) was added and reaction was leftstirring for 90 minutes. Reaction mixture was concentrated in vacuo anddried on vacuum pump overnight to yield intermediate5-amino-3-(3-fluoro-5-methoxyphenyl)-1H-pyrazole-4-carbonitrile (BA73).Formamide (20 ml) was added and reaction was heated to 180° C. under anargon atmosphere overnight. Reaction was cooled and dH2O was added (40ml) forcing a white precipitate out of solution. Precipitate wascollected and washed with dH2O. Solid was dried and purified by silicagel column chromatography [MeOH—CH2Cl2, 10:90] to yield3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA75).

3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA75,100 mg, 0.386 mmol) was dissolved in DMF (10 ml). K2CO3 (250 mg, 1.54mmol) was added and reaction was stirred at room temperature under anargon atmosphere. lodocyclopentane (0.134 ml, 1.16 mmol) was added witha syringe and reaction was stirred for 2 hours. Solid K2CO3 was removedby filtration. Solvent was partially removed in vacuo. Sodium citrate(50 ml) was added and reaction was extracted with EtOAc. Organic phasesconcentrated in vacuo and purified using by RP-HPLC (MeCN:H2O:0.1% TFA)to yield BA79.

Synthesis of1-cyclopentyl-3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA79d).1-cyclopentyl-3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA79, 0.386 mmol) was dissolved in CH2Cl2 (2 ml). BBr3 (4 mL, 4 mol)was added slowly with a syringe, while stirring. The reaction wasstirred at room temperature for 2 hours then concentrated in vacuo andpurified using by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA79 (69 mg, 57%yield).

Synthesis of 1-cyclopentyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA80). A solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (400mg, 1.53 mmol) and K2CO3 (1 g, 6 mmol) in DMF (5 ml) was stirred at roomtemperature under an argon atomosphere. Iodocyclopentane (1.0 g, 0.0084mol) was added with a syringe. Reaction was refluxed under argonatmosphere for 2 hours. Solid K2CO3 was removed by filtration. Solventwas partially removed in vacuo. Sodium citrate (50 ml) was added andreaction was extracted with EtOAc. Organic phases concentrated in vacuoand purified using silica gel column chromatography [MeOH—CH2Cl2, 5:95]yielding BA80 (300 mg, 60% yield). ESI-MS (M+H)+ m/z calcd 330.0, found330.0.

Synthesis of1-(3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)ethanone(BA81, BA81d & BA81dd). A solution of tert-butyl2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate(200 mg, 0.76 mmol) in EtOH (3.3 ml) was added to a solution of1-cyclopentyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA80, 100 mg,0.30 mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturatedNa2CO3 (1.9 ml) were added and the reaction was heated to 80° C. underan argon atmosphere overnight. After cooling, the reaction was extractedwith saturated NaCl and CH2Cl2. Organic phases were combined,concentrated in vacuo and purified using silica gel columnchromatography [MeOH—CH2Cl2, 5:95] yielding BA81. BA81 was dissolved in50:50 CH2Cl2:TFA and stirred for one hour at room temperature. Thereaction mixture was concentrated in vacuo and purified using by RP-HPLC(MeCN:H2O:0.1% TFA) to yield BA81d. BA81d was dissolved in CH2Cl2 (2 ml)and BBr3 (4 mL, 4 mol) was added slowly with a syringe, while stirring.The reaction was stirred at room temperature for 2 hours thenconcentrated in vacuo and purified using by RP-HPLC (MeCN:H2O:0.1% TFA)to yield BA81dd.

Synthesis of3-(3-bromo-5-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA85). A solution of2-(3-bromo-5-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (137mg, 0.43 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (65 mg, 0.216mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA85 (28 mg,36% yield). ESI-MS (M+H)+ m/z calcd 362.1, found 362.0.

Synthesis of3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5-bromophenol(BA87).3-(3-bromo-5-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA85, 0.1 mmol) was dissolved in CH2Cl2 (1 ml) and BBr3 (1 mL, 1 mol)was added slowly with a syringe, while stirring. The reaction wasstirred at room temperature for 35 minutes then concentrated in vacuoand purified using by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA87 (10.7mg, 31% yield). ESI-MS (M+H)+ m/z calcd 348.0, found 348.0.

Synthesis of tert-butyl5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-1-carboxylate(BA86). A solution of tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylate(212 mg, 0.61 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (75 mg, 0.25mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O) to yield BA86 (9.3 mg, 9%yield). EST-MS (M+H)+ m/z calcd 362.1, found 362.0.

Synthesis of3-(1H-indol-5-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA89). Tert-butyl5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-1-carboxylate(BA86, 9 mg, 0.022 mmol) was dissolved in 50:50 CH2Cl2:TFA and stirredfor one hour at room temperature. The reaction mixture was concentratedin vacuo and purified using by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA89(4.8 mg, 75% yield). ESI-MS (M+H)+ m/z calcd 293.1, found 293.0.

Synthesis of tert-butyl5-(4-amino-1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-1-carboxylate(BA88). A solution of tert-butyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylate(130 mg, 0.38 mmol) in EtOH (3.3 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg, 0.15mmol) in DME (12 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(1.9 ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O) to yield BA88. ESI-MS (M+H)+m/z calcd 419.2, found 419.1.

Synthesis of3-(1H-indol-5-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA94). Tert-butyl5-(4-amino-1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indole-1-carboxylate(BA88) was dissolved in 50:50 CH2Cl2:TFA and stirred for one hour atroom temperature. The reaction mixture was concentrated in vacuo andpurified using by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA89 (6.3 mg).ESI-MS (M+H)+ m/z calcd 319.1, found 319.2.

Synthesis of1-cyclopentyl-3-(3,4-dimethoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA90). A solution of 3,4-dimethoxyphenylboronic acid (41 mg, 0.23.mmol) in EtOH (1.65 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.09mmol) in DME (6 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(0.95 ml) were added and the reaction was heated to 80° C. under anargon atmosphere overnight. After cooling, the reaction was extractedwith saturated NaCl and CH2Cl2. Organic phases were combined,concentrated in vacuo and purified by RP-HPLC (MeCN:H2O) to yield BA90(8.4 mg, 28% yield). ESI-MS (M+H)+ m/z calcd 340.2, found 340.1.

Synthesis of3-(1H-indol-4-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA91). A solution of 1H-indol-4-yl-4-boronic acid (40 mg, 0.25 mmol) inEtOH (1.65 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.1 mmol)in DME (6 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3 (0.95ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA91 (14.6mg, 50% yield). ESI-MS (M+H)+ m/z calcd 293.1, found 293.1.

Synthesis of1-cyclopentyl-3-(1H-indol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA92). A solution of 1H-indol-4-yl-4-boronic acid (30 mg, 0.19 mmol) inEtOH (1.65 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (25 mg, 0.076mmol) in DME (6 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3(0.95 ml) were added and the reaction was heated to 80° C. under anargon atmosphere overnight. After cooling, the reaction was extractedwith saturated NaCl and CH2Cl2. Organic phases were combined,concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) toyield BA92 (23 mg, 95% yield). ESI-MS (M+H)+ m/z calcd 319.2, found319.1.

Synthesis of3-(2,3-dihydrobenzofuran-5-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA95). A solution of 2,3-dihydro-1-benzofaran-5-ylboronic acid (38 mg,0.23 mmol) in EtOH (1.65 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.1 mmol)in DME (6 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3 (0.95ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA95 (15.7mg, 59% yield). ESI-MS (M+H)+ m/z calcd 296.1, found 296.1.

Synthesis of3-(benzofuran-5-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA96). A solution of5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-benzofuran (56 mg,0.23 mmol) in EtOH (1.65 ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (30 mg, 0.1 mmol)in DME (6 ml). Pd(PPh3)4 (30 mg, 0.03 mmol) and saturated Na2CO3 (0.95ml) were added and the reaction was heated to 80° C. under an argonatmosphere overnight. After cooling, the reaction was extracted withsaturated NaCl and CH2Cl2. Organic phases were combined, concentrated invacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA96 (19 mg,72% yield). ESI-MS (M+H)+ m/z calcd 296.1, found 296.1.

Synthesis of5-(4-amino-1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-ethoxyphenol(BA98).1-cyclopentyl-3-(4-ethoxy-3-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK359, 25 mg, 0,071 mmol) was dissolved in CH2Cl2 (5 ml) and stirred at−10° C. under an argon atmosphere. After 30 minutes, reaction wasbrought to 0° C. and stirred for 2.5 hours. Reaction was stirred foradditional 4 hours at room temperature, then concentrated in vacuo andpurified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA98 (3 mg, 13% yield).ESI-MS (M+H)+ m/z calcd 340.1, found 340.1.

Synthesis of2-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenylamino)propan-1-ol(BA99).3-(4-amino-3-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA20d) (30 mg, 0.10 mmol) was dissolved in DMF (0.400 ml). K2CO3 (55mg, 0.4 mmol) was added and reaction was stirred at 70° C.3-bromo-1-propanol (0.050 ml, 0.6 mmol) was added and reaction wasstirred overnight. Solid K2CO3 was removed by filtration. Solvent waspartially removed in vacuo. Sodium citrate (50 ml) was added andreaction was extracted with saturated NaCl and CH2Cl2. Organic phasesconcentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) toyield BA99 (8.4 mg, 24% yield). ESI-MS (M+H)+ m/z calcd 357.2, found357.1.

Synthesis of 3-iodo-1-methyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA109). A solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2 g,7.69 mmol) and K2CO3 (4.25 g, 30.8 mmol) in DMF (5 ml) was stirred atroom temperature under an argon atomosphere. Iodomethane (1.17 ml, 7.69mmol) was added with a syringe. Reaction was stirred under an argonatmosphere at room temperature for 2 hours. Solid K2CO3 was removed byfiltration. Solvent was partially removed in vacuo. Sodium citrate (50ml) was added and reaction was extracted with EtOAc. Organic phasesconcentrated in vacuo and purified using silica gel columnchromatography [MeOH—CH2Cl2, 5:95] yielding BA109 (212 mg, 10% yield).ESI-MS (M+H)+ m/z calcd 275.9, found 275.9

General synthetic scheme for BA102, BA105-108, BA110, BA118,BA128-BA135, BA137, BA139-140, BA143, BA147, BA149-BA152, BA156, BA158,BA160-BA162. A solution of boronic acid (2.5 equivalents) in EtOH (1.65ml) was added to a solution of3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA12, 1equivalent) in DME (6 ml). Pd(PPh3)4 (15 mg, 0.15 mmol) and saturatedNa2CO3 (0.95 ml) were added and the reaction was heated to 90° C. underan argon atmosphere overnight. After cooling, the reaction was extractedwith saturated NaCl and CH2Cl2. Organic phases were combined,concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) toyield desired products. Products were analyzed by LC-MS.

General synthetic scheme for BA112, BA115, BA121, BA122, BA124, BA136,BA138. BA141, and BA144. A solution of boronic acid (2.5 equivalents) inEtOH (1.65 ml) was added to a solution of1-cyclopentyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA80, 1equivalent) in DME (6 ml). Pd(PPh3)4 (15 mg, 0.15 mmol) and saturatedNa2CO3 (0.95 ml) were added and the reaction was heated to 90° C. underan argon atmosphere overnight. After cooling, the reaction was extractedwith saturated NaCl and CH2Cl2. Organic phases were combined,concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) toyield desired products. Products were analyzed by LC-MS.

General synthetic scheme for BA111, BA114, BA116, BA117, BA119, andBA120. A solution of boronic acid (2.5 equivalents) in EtOH (1.65 ml)was added to a solution of3-iodo-1-methyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (BA109, 1equivalent) in DME (6 ml). Pd(PPh3)4 (15 mg, 0.15 mmol) and saturatedNa2CO3 (0.95 ml) were added and the reaction was heated to 90° C. underan argon atmosphere overnight. After cooling, the reaction was extractedwith saturated NaCl and CH2Cl2. Organic phases were combined,concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1% TFA) toyield desired products. Products were analyzed by LC-MS.

Synthesis of6-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)quinolin-2-amine(BA146).3-(2-chloroquinolin-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA130, 50 mg, 0.15 mmol), acetamide (174 mg, 3.0 mmol) and K2CO3 (104mg, 0.75 mmol) were combined and heated to 200° C. under an argonatmosphere for one hour. Reaction was cooled, then extracted with H2Oand CH2Cl2. Organic phases were combined, concentrated in vacuo andpurified by RP-HPLC (MeCN:H2O:0.1% TFA) to yield BA146 (22 mg, 46%yield). ESI-MS (M+H)+ m/z calcd 320.2, found 320.4.

Synthesis of3-(3-amino-1H-indazol-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA154).4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorobenzonitrile(BA150, 20 mg, 0.07 mmol) was dissolved in n-BuOH (2 ml). Hydrazinemonohydrate (0.400 ml) was added and the reaction was heated to 110° C.under an argon atomosphere and left stirring over night. Reactionmixture was concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1%TFA) to yield BA154 (15 mg, 70% yield). ESI-MS (M+H)+ m/z calcd 309.2,found 309.4.

Synthesis of4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-hydroxybenzonitrile(BA155_(—)2).4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorobenzonitrile(BA150, 25 mg, 0.1 mmol) was dissolved in DMF (1 ml). t-BuOK (24 mg,0.21 mmol) was added and the reaction was stirred at room temperatureovernight. Reaction was then heated to 150° C. for 24 hours. Thereaction was then concentrated in vacuo and purified by RP-HPLC(MeCN:H2O:0.1% TFA) to yield BA155_(—)2 (21 mg, 89% yield). ESI-MS(M+H)+ m/z calcd 295.1, found 295.4.

Synthesis of3-(3-aminobenzo[d]isoxazol-5-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA157_(—)2) &5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-hydroxybenzonitrile(BA157_(—)3).5-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorobenzonitrile(BA151, 20 mg, 0.07 mmol) was dissolved in DMF (1 ml). t-BuOK (24 mg,0.21 mmol) was added and the reaction was stirred at room temperatureovernight. Reaction was then heated to 150° C. for 24 hours. Thereaction was then concentrated in vacuo and purified by RP-HPLC(MeCN:H2O:0.1% TFA) to yield BA157_(—)2 (7 mg), ESI-MS (M+H)+ m/z calcd295.1, found 295.4 and BA157_(—)3 (8 mg), ESI-MS (M+H)+ m/z calcd 310.1,found 310.4.

Synthesis of3-(3-amino-1H-indazol-6-yl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(BA159).4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2,6-difluorobenzaldehyde(BA149, 20 mg, 0.063 mmol) was dissolved in n-BuOH (1 ml). Hydrazinemonohydrate (0.400 ml) was added and the reaction was heated to 100° C.under an argon atomosphere and left stirring for 2.5 hours. Reactionmixture was concentrated in vacuo and purified by RP-HPLC (MeCN:H2O:0.1%TFA) to yield BA159 (15 mg, 77% yield). ESI-MS (M+H)+ m/z calcd 312.1,found 312.4.

Synthesis of4-chloro-7-methyl-5-(naphthalen-2-yl)-7H-pyrrolo[2,3-d]pyrimidine(ZK102). A solution of4-chloro-5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (19 mg, 0.065mmol), naphthalen-2-yl-2-boronic acid (12.2 mg, 0.071 mmol), Na₂CO₃(68.9 mg, 0.65 mmol) and PdCl₂(dppf) (26.5 mg, 0.00325 mmol) in THF (3mL) was heated to reflux overnight under an argon atmosphere. Reactionwas concentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) toyield ZK102 (5 mg, 26% yield). ESI-MS (M+H)⁺ m/z calcd 294.1, found294.3.

Synthesis of4-chloro-7-methyl-5-(3-biphenyl)-7H-pyrrolo[2,3-d]pyrimidine (ZK103). Asolution of 4-chloro-5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (10 mg,0.034 mmol), 3-biphenyl-boronic acid (7.4 mg, 0.038 mmol), Na₂CO₃ (36.1mg, 0.34 mmol) and PdCl₂(dppf) (1.4 mg, 0.0017 mmol) in THF (10 mL) washeated to reflux overnight under an argon atmosphere. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA) toyield ZK103 (3 mg, 28% yield). ESI-MS (M+H)⁺ m/z calcd 320.1, found320.0.

Synthesis of 3-(4-tert-butylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK125).1-tert-butyl-3-(4-tert-butylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-aminewas dissolved in a solution of formic acid (1 mL) and conc. HCl (0.1 mL)and heated to reflux for 2 hours. Reaction was concentrated in vacuo andpurified by RP-HPLC (MeCN:H₂O:0.1% TFA) to yield ZK125 (quant.). ESI-MS(M+H)⁺ m/z calcd 268.2, found 268.4.

Synthesis of 3-(3-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK126).1-tert-butyl-3-(3-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50mg, 0.16 mmol) was dissolved in a solution of formic acid (5 mL) andconc. HCl (0.1 mL) and heated to reflux for 2.5 hours. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 304.1, found 304.3.

Synthesis of 3-m-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (ZK127).1-tert-butyl-3-m-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (23 mg, 0.1mmol) was dissolved in a solution of formic acid (1 mL) and conc. HCl(0.3 mL) and heated to reflux for 2.5 hours. Reaction was concentratedin vacuo and purified-by RP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/zcalcd 226.1, found 226.3.

Synthesis of 3-(3-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK128).1-tert-butyl-3-(3-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (23mg, 0.055 mmol) was dissolved in a solution of formic acid (5 mL) andconc. HCl (0.1 mL) and heated to reflux for 2 hours. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 257.1, found 257.3.

Synthesis of3-(benzo[d][1,3]dioxol-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK129).1-tert-butyl-3-(benzo[d][1,3]dioxol-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(21 mg, 0.082 mmol) was dissolved in a solution of formic acid (1 mL)and conc. HCl (0.2 mL) and heated to reflux for 2 hours. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 256.1, found 256.3.

Synthesis of 3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK130).1-tert-butyl-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (21mg, 0.082 mmol) was dissolved in a solution of formic acid (2 mL) andconc. HCl (0.2 mL) and heated to reflux for 30 min. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 257.1, found 257.3.

Synthesis of3-(3-(2,6-dichlorobenzyloxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK131).1-tert-butyl-3-(3-(2,6-dichlorobenzyloxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(19.5 mg, 0.05 mmol) was dissolved in a solution of formic acid (2 mL)and conc. HCl (0.2 mL) and heated to reflux for 30 min. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 386.1, found 386.2.

Synthesis of 3-(2,3-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK132).1-tert-butyl-3-(2,3-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(34 mg, 0.14 mmol) was dissolved in a solution of formic acid (2 mL) andconc. HCl (0.2 mL) and heated to reflux for 30 min. Reaction wasconcentrated in vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 240.1, found 240.4.

Synthesis of 2-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol (ZK133).1-tert-butyl-2-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol (5 mg,0.014 mmol) was dissolved in a solution of formic acid (2 mL) and conc.HCl (0.2 mL) and heated to reflux for 30 min. Reaction was concentratedin vacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/zcalcd 228.1, found 228.3.

Synthesis of 3-o-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (ZK134).1-tert-butyl-3-o-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine was dissolvedin a solution of formic acid (2 mL) and conc. HCl (0.2 mL) and heated toreflux for 30 min. Reaction was concentrated in vacuo and purified byRP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 226.1, found 226.3.

Synthesis of 3-(3-aminophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK135).1-tert-butyl-3-(3-aminophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine wasdissolved in a solution of formic acid (2 mL) and conc. HCl (0.2 mL) andheated to reflux for 30 min. Reaction was concentrated in vacuo andpurified by RP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 227.1,found 227.3.

Synthesis of 3-(3-(benzyloxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK136).1-tert-butyl-3-(3-(benzyloxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(19 mg, 0.052 mmol) was dissolved in a solution of formic acid (1 mL)and conc. HCl (0.1 mL) and heated to reflux for 30 min. Reaction yieldeda mixture of ZK136 and3-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenol (ZK138). Reaction wasconcentrated in vacuo and the products purified by RP-HPLC(MeCN:H₂O:0.1% TFA). ZK136: ESI-MS (M+H)⁺ m/z calcd 318.1, found 318.3.ZK138: ESI-MS (M+H)⁺ m/z calcd 228.1, found 228.3.

Synthesis of 3-(4-aminophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK137).1-tert-butyl-3-(4-aminophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (9mg, 0.032 mmol) was dissolved in a solution of formic acid (1 mL) andconc. HCl (0.2 mL) and heated to reflux. The reaction was allowed toproceed 30 min., then concentrated in vacuo and the products purified byRP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 227.1, found 227.3.

Synthesis of3-(1,2,3,4-tetrahydronaphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK139).1-tert-butyl-3-(1,2,3,4-tetrahydronaphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(9 mg, 0.029 mmol) was dissolved in a solution of formic acid (1 mL) andconc. HCl (0.1 mL) and heated to reflux. The reaction was allowed toproceed 30 min., then concentrated in vacuo and the products purified byRP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 266.1, found 266.4.

Synthesis of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (ZK140).4-chloro-5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (90 mg, 0.31 mmol)was taken up in 7N NH₃/MeOH and heated in a sealed tube at 110° C.overnight. Reaction was concentrated in vacuo to give a brown/off-whitesolid.

Synthesis of 3-p-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (ZK141).1-tert-butyl-3-p-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine was dissolvedin a solution of formic acid (1 mL) and conc. HCl (0.1 mL) and heated toreflux. The reaction was allowed to proceed 30 min., then concentratedin vacuo and the products purified by RP-HPLC (MeCN:H₂O:0.1% TFA).ESI-MS (M+H)⁺ m/z calcd 226.1, found 226.3.

Synthesis of 3-(4-biphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (ZK142).1-tert-butyl-3-(4-biphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (22 mg,0.066 mmol) was dissolved in a solution of formic acid (1 mL) and conc.HCl (0.1 mL) and heated to reflux. The reaction was allowed to proceed30 min., then concentrated in vacuo and the products purified by RP-HPLC(MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 288.1, found 288.3.

Synthesis of 3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK143).1-tert-butyl-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (19mg, 54.1 mmol) was dissolved in a solution of formic acid (1 mL) andconc. HCl (0.1 mL) and heated to reflux. The reaction was allowed toproceed 30 min., then concentrated in vacuo and the products purified byRP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 304.1, found 304.3.

Synthesis of1-benzyl-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK147). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (110mg, 0.42 mmol) was dissolved in DMF (2 mL) and K₂CO₃ (220 mg, 1.6 mmol)and benzyl bromide (71.8 mg, 0.42 mmol) were added. The reaction washeated to 60° C. overnight, then cooled to RT and poured into water (30mL). The precipitate was collected by filtration and then purifiedfurther by silica gel chromatography (5% MeOH/CH₂Cl₂) to yield a whitesolid.

Synthesis of5-(4-(benzyloxy)phenyl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine(ZK150). A solution of5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (5 mg, 0.018 mmol),4-(benzyloxy)phenylboronic acid (21 mg, 0.091 mmol), K₃PO₄ (19.3 mg,0.091 mmol) and Pd(PPh₃)₄ (12.5 mg, 0.011 mmol) in DMF (3 mL) was heatedto 60° C. under an argon atmosphere. Reaction was concentrated in vacuoand purified by RP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd331.1, found 331.3.

Synthesis of 5-(3-biphenyl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine(ZK151). A solution of5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (5 mg, 0.018 mmol),3-biphenylboronic acid (18 mg, 0.091 mmol), K₃PO₄ (19.3 mg, 0.091 mmol)and Pd(PPh₃)₄ (12.5 mg, 0.011 mmol) in DMF (3 mL) was heated to 60° C.under an argon atmosphere. Reaction was concentrated in vacuo andpurified by RP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/z calcd 301.1,found 301.3.

Synthesis of5-(benzo[b]thiophen-2-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine(ZK152). A solution of5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (5 mg, 0.018 mmol),benzo[b]thiophen-2-yl-2-boronic acid (16 mg, 0.091 mmol), K₃PO₄ (19.3mg, 0.091 mmol) and Pd(PPh₃)₄ (12.5 mg, 0.011 mmol) in DMF (3 mL) washeated to 60° C. under an argon atmosphere. Reaction was concentrated invacuo and purified by RP-HPLC (MeCN:H₂O:0.1% TFA). ESI-MS (M+H)⁺ m/zcalcd 281.1, found 281.3.

Synthesis of3-(naphthalen-2-yl)-1-phenethyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK155). 2-(methoxy(naphthalen-6-yl)methylene)malononitrile (100 mg,0.43 mmol) and phenethylhydrazine hydrogen chloride (58.5 mg, 0.43 mmol)were dissolved in EtOH (3 mL) and TEA (60 μL, 0.43 mmol) and heat toreflux for one hour. The product was extracted with diethylether andconcentrated in vacuo. This concentrate was then dissolved in formamide(10 mL) and heated to 160-180° C. overnight. The following day thereaction was cooled, poured into water, and the precipitated productcollected by filtration. ESI-MS (M+H)⁺ m/z calcd 366.2, found 366.2.

Synthesis of1-isopropyl-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK156). 2-(methoxy(naphthalen-6-yl)methylene)malononitrile (100 mg,0.43 mmol) and isopropylhydrazine hydrogen chloride (47.3 mg, 0.43 mmol)were dissolved in EtOH (3 mL) and TEA (1 eq.) and heat to reflux for onehour. The product was extracted with diethylether and concentrated invacuo. This concentrate was then dissolved in formamide (10 mL) andheated to 160-180° C. overnight. The following day the reaction wascooled, poured into water, and the precipitated product collected byfiltration. ESI-MS (M+H)⁺ m/z calcd 304.2, found 304.2.

Synthesis of1-ethyl-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (ZK157).3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (100 mg, 0.42mmol) was dissolved in DMF (3 mL) and K₂CO₃ (220 mg, 1.6 mmol) and ethyliodide (37 μL, 0.46 mmol) were added. The reaction was heated to 60° C.overnight, then cooled to RT and poured into water (30 mL). Theprecipitate was collected by filtration. ESI-MS (M+H)⁺ m/z calcd 290.1,found 290.2.

Synthesis of1-cyclopentyl-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK158). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (100mg, 0.42 mmol) was dissolved in DMF (3 mL) and K₂CO₃ (220 mg, 1.6 mmol)and cyclopentyl bromide (49.5 μL, 0.46 mmol) were added. The reactionwas heated to 60° C. overnight, then cooled to RT and poured into water(30 mL). The precipitate was collected by filtration. ESI-MS (M+H)⁺ m/zcalcd 330.2, found 330.2.

Synthesis of1-allyl-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (ZK159).3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg, 0.21mmol) was dissolved in DMF (1.5 mL) and K₂CO₃ (110 mg, 0.8 mmol) andallyl iodide (23 μL, 0.25 mmol) were added. The reaction was heated to60° C. overnight, then cooled to RT and poured into water (30 mL). Theprecipitate was collected by filtration. ESI-MS (M+H)⁺ m/z calcd 302.1,found 302.2.

Synthesis of2-(4-amino-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)acetamide(ZK162). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg,0.21 mmol) was dissolved in DMF (1.5 mL) and K₂CO₃ (110 mg, 0.8 mmol)and iodoacetamide (46 mg, 0.25 mmol) were added. The reaction was heatedto 60° C. overnight, then cooled to RT and poured into water (30 mL).The precipitate was collected by filtration. ESI-MS (M+H)⁺ m/z calcd319.1, found 319.2.

Synthesis of1-(cyclopropylmethyl)-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK165). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg,0.21 mmol) was dissolved in DMF (1.5 mL) and K₂CO₃ (110 mg, 0.8 mmol)and cyclopropyl methyl bromide (22 μL, 0.25 mmol) were added. Thereaction was heated to 60° C. overnight, then cooled to RT and pouredinto water (30 mL). The precipitate was collected by filtration. ESI-MS(M+H)⁺ m/z calcd 316.2, found 316.2.

Synthesis of1-isopentyl-3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK161). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg,0.21 mmol) was dissolved in DMF (1.5 mL) and K₂CO₃ (110 mg, 0.8 mmol)and isobutyl bromide were added. The reaction was heated to 60° C.overnight, then cooled to RT and poured into water (30 mL). Theprecipitate was collected by filtration. ESI-MS (M+H)⁺ m/z calcd 332.2,found 332.3.

Synthesis of3-(naphthalen-2-yl)-1-((E)-3-phenylprop-1-enyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK167). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg,0.21 mmol) was dissolved in DMF (1.5 mL) and K₂CO₃ (110 mg, 0.8 mmol)and 1-((E)-3-bromoprop-1-enyl)benzene were added. The reaction washeated to 60° C. overnight, then cooled to RT and poured into water (30mL). The precipitate was collected by filtration. ESI-MS (M+H)⁺ m/zcalcd 378.2, found 378.2.

Synthesis of3-(naphthalen-2-yl)-1-(prop-2-ynyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK168). 3-(naphthalen-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50 mg,0.21 mmol) was dissolved in DMF (1.5 mL) and K₂CO₃ (110 mg, 0.8 mmol)and propargylbromide were added. The reaction was heated to 60° C.overnight, then cooled to RT and poured into water (30 mL). Theprecipitate was collected by filtration. ESI-MS (M+H)⁺ m/z calcd 300.1,found 300.2

Synthesis of 3-ethoxy-4-methoxybenzoyl chloride (ZK299).3-ethoxy-4-methoxybenzoic acid (5 g, 25.5 mmol) was added to a solutionof CH₂Cl₂ (40 mL) and benzene (20 mL) in a flame-dried 150 mL roundbottom flask. Anhydrous DMF (9 drops) was added and the solution wascooled on ice. Oxalyl chloride (11 mL, 128 mmol) was added dropwise, andthe reaction was then allowed to warm to RT. The reaction was stirred atRT for 90 minutes, then concentrated in vacuo yield an off-white solid.The solid was placed on a high-vacuum line for 2 hours, and then takenonto the next step without further characterization.

Synthesis of2-((3-ethoxy-4-methoxyphenyl)(hydroxy)methylene)malononitrile (ZK301).NaH (2.2 g, 56 mmol, 60% dispersion in paraffin oil) was added to asolution of malononitrile (1.85 g, 28 mmol) in THF (30 mL) on ice.3-ethoxy-4-methoxybenzoyl chloride (25.5 mmol) was dissolved in THF (50mL) and added the first solution dropwise by syringe at 0° C. The icewas then removed and the reaction was allowed to proceed at RT for 60min. 1N HCl (100 mL) was added and the solution was extracted threetimes with EtOAc. The organic phase was dried with MgSO₄, filtered, andconcentrated in vacuo to give an orange solid that was taken onto thenext step without further characterization.

Synthesis of2-((3-ethoxy-4-methoxyphenyl)(methoxy)methylene)malononitrile (ZK302).2-((3-ethoxy-4-methoxyphenyl)(hydroxy)methylene)malononitrile (25.5mmol) and sodium bicarbonate (17 g, 204 mmol) were combined in asolution of 1,4-dioxane (48 mL) and water (8 mL). Dimethylsulphate (17mL, 178 mmol) was slowly added and the reaction was heated to 80-90° C.for 2 hours. The reaction was cooled to RT, water was added, and theaqueous phase extracted three times with EtOAc (200 mL). The organicphases were combined, dried with MgSO₄, and filtered to give a red oil.The oil was purified by silica gel chromatography (10% EtOAc/Hexanes,R_(f)˜0.1) to give a white solid (3.59 g, 54.5% yield over three steps).ESI-MS (M+H)⁺ m/z calcd 259.1, found 259.0.

Synthesis of5-amino-3-(3-ethoxy-4-methoxyphenyl)-1-isopropyl-1H-pyrazole-4-carbonitrile(ZK303). 2-((3-ethoxy-4-methoxyphenyl)(methoxy)methylene)malononitrile(200 mg, 0.78 mmol), isopropylhydrazine hydrogen chloride (86 mg, 0.78mmol), and triethylamine (0.10 mL, 0.78 mmol) were combined in ethanol(5 mL) and heated to reflux for 90 minutes. The reaction was then cooledto RT, water was added and the aqueous phase was extracted three timeswith EtOAc. The organic phase was concentrated and carried onto the nextstep without further characterization. ESI-MS (M+H)⁺ m/z calcd 301.1,found 301.0

Synthesis of3-(3-ethoxy-4-methoxyphenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine(ZK305).5-amino-3-(3-ethoxy-4-methoxyphenyl)-1-isopropyl-1H-pyrazole-4-carbonitrilewas dissolved in formamide (20 mL) and heated to 180° C. overnight. Thenext day the reaction was cooled to RT, water was added, and theprecipitate was collected by filtration. The precipitate was thendissolved in CH₂Cl₂/MeOH and passed through a silica plug. The productwas then lyophilized from benzene to yield an off-white solid (48 mg,19% over two steps). ESI-MS (M+H)⁺ m/z calcd 328.2, found 328.0.

Synthesis of5-amino-3-(3-ethoxy-4-methoxyphenyl)-1-(2-hydroxyethyl)-1H-pyrazole-4-carbonitrile(ZK304). 2-((3-ethoxy-4-methoxyphenyl)(methoxy)methylene)malononitrile(200 mg, 0.78 mmol), 2-hydroxyethylhydrazine (0.056 mL, 0.78 mmol), andtriethylamine (0.10 mL, 0.78 mmol) were combined in ethanol (5 mL) andheated to reflux for 90 minutes. The reaction was then cooled to RT,water was added and the aqueous phase was extracted three times withEtOAc, CH₂Cl₂, and CHCl₃. The organic phase was concentrated and carriedonto the next step without further characterization. ESI-MS (M+H)⁺ m/zcalcd 303.1, found 303.0.

2-(4-amino-3-(3-ethoxy-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethanol(ZK306).5-amino-3-(3-ethoxy-4-methoxyphenyl)-1-(2-hydroxyethyl)-1H-pyrazole-4-carbonitrilewas dissolved in formamide (20 mL) and heated to 180° C. overnight. Thenext day the reaction was cooled to RT, water was added, and theprecipitate was collected by filtration. The precipitate was thendissolved in CH₂Cl₂/MeOH and passed through a silica plug. The productwas then lyophilized from benzene to yield a brown solid (6.4 mg, 2.5%over two steps). ESI-MS (M+H)⁺ m/z calcd 330.1, found 330.0.

B. Structural Studies

Crystal structures of p110γ have been reported, alone and in complexwith ATP or pan-specific inhibitors such as LY294002 and wortmannin(Walker et al., 2000; Walker et al., 1999). To explore how potent andselective inhibitors bind, the crystal structures of PI3-K inhibitorsfrom three chemotypes bound to human p110γ were determined at 2.5-2.6 Åresolution: the quinazoline purine PIK-39, the imidazopyridine PIK-90and the phenylthiazole PIK-93 (FIG. 2).

Based on these co-crystal structures and a conserved arylmorpholinepharmacophore model, structural models were generated for threeadditional chemotypes bound to p110γ: the pyridinylfuranopyrimidinePI-103, the morpholinochromone PIK-108, and the morpholinopyranoneKU-55933 (FIG. 2). Model-building for these inhibitors was guided by theobservation that each compound contains the key arylmorpholinepharmacophore found in LY294002.

PIK-39 is an isoquinoline purine that inhibits p110δ at mid-nanomolarconcentrations, p110γ and p110β at concentrations ˜100-fold higher, andshows no activity against any other PI3-K family member, includingp110α, at concentrations up to 100 μM (FIG. 5). The biochemicalselectivity of this compound is achieved through an unusual binding moderevealed in its co-crystal structure with p110γ (FIG. 2C). Only themercaptopurine moiety of PIK-39 makes contacts within the interior ofthe ATP binding pocket, and this ring system is rotated ˜110° andtwisted ˜35° out of the plane relative to the adenine of the ATP. Inthis orientation, it satisfies hydrogen bonds to the backbone amides ofVal 882 and Glu 880 (thereby recapitulating the hydrogen bonds made byN1 and N6 of adenine).

In contrast to other PI3-K inhibitor structures, PIK-39 does not accessthe deeper pocket in the active site interior (FIG. 2C, lightly shadedarea labeled as “Affinity Pocket”). Instead, the aryl-isoquinolinemoiety of PIK-39 extends out to the entrance of the ATP binding pocket(FIG. 2B). In this region, the kinase accommodates the inhibitor byundergoing a conformational rearrangement in which Met 804 shifts froman “up” position, in which it forms the ceiling of the ATP bindingpocket, to a “down” position which it packs against the isoquinolinemoiety. The effect of this movement, which is unique to the PIK-39structure (FIG. 2B), is to create a novel hydrophobic pocket between Met804 and Trp 812 at the entrance to the ATP binding site. Thisinduced-fit pocket buries ˜180 Å2 of solvent accessible inhibitorsurface area, enabling PIK-39 to achieve nanomolar affinity despitelimited contacts within the active site core.

Co-crystal structures of PIK-90 and PIK-93 compounds bound to p110γ weredetermined. PIK-90 and PIK-93 both make a hydrogen bond to the backboneamide nitrogen of Val 882 (FIG. 2D), an interaction conserved among allknown PI3-K inhibitors (Walker et al., 2000). In addition to thishydrogen bond, PIK-93 makes a second hydrogen bond to the backbonecarbonyl of Val 882 and a third between its sulphonamide moiety and theside chain of Asp 964. PIK-93 is one of the most polar inhibitors in ourpanel (clogP=1.69) and these extended polar interactions may compensatefor its limited hydrophobic surface area.

PIK-90 binds in a mode similar to PIK-93, although this larger compoundmakes more extensive hydrophobic interactions, burying 327 Å² of solventaccessible surface area. To achieve this, PIK-90 projects its pyridinering into a deeper cavity that is partially accessed by PIK-93 but notoccupied by ATP (FIG. 2D, lightly shaded circle). In this region, thepyridine ring of PIK-90 is poised to make a hydrogen bond to Lys 833,and we find that replacement of this pyridine nitrogen with carbonresults in a 100-fold loss in affinity (PIK-95, FIG. 4). PI-103, a thirdmulti-targeted PI3K inhibitor, projects a phenol into the same pocketbased on an arylmorpholine pharmacophore model (FIG. 2D).

Two structural features distinguish these potent, multi-targetedinhibitors from the more selective compounds in our panel. First, thesecompounds adopt a flat conformation in the ATP binding pocket, whereashighly selective inhibitors project out of the plane occupied by ATP(FIG. 2). Second, the most potent inhibitors project into a deeperbinding pocket that is not accessed by ATP (FIG. 2A). Much of thesurface of this affinity pocket is contributed by the side-chain of Ile879.

The mercaptopurine in the PIK-39 structure was replaced with adenine toyield a model of IC87114 (FIG. 3A). This substitution provided theadenine of IC87114 in the correct orientation to make the same hydrogenbonds as the mercaptopurine of PIK-39, even though these two ringsystems are rotated by 110° with respect to each other.

Unlike other inhibitor chemotypes, PIK-39 does not exploit thePI3-kinase affinity pocket (FIG. 2C). The pyrazolopyrimidine analog ofIC87114 (PIK-293) as well as a novel analog containing an m-phenol(PIK-294, FIG. 3A) were then tested for inhibition of the class IPI3-Ks. PIK-294 was up to 60-fold more potent than PIK-293 (FIG. 3A).

The structure of PIK-39 bound to p110γ reveals a conformationalrearrangement of Met 804 that creates an induced pocket, and we havehypothesized that this conformational rearrangement underlies theselectivity of PIK-39 for p110δ. A prediction of this model is thatmutation of Met 804 should perturb the binding of p110δ-selectiveinhibitors (which access the induced pocket), but not affect otherclasses of inhibitors (which do not access this pocket). Modelingsuggests that mutation of Met 804 to a δ-branched amino acid (such asvaline or isoleucine) should restrict the pocket formed by rearrangementof that residue (FIG. 3B, right). Therefore, we mutated thecorresponding residue in p110δ (Met 752) to valine or isoleucine,expressed and purified these kinases, and tested them for sensitivity toPI3-K inhibitors (FIG. 3B). We find that M752I and M752V p110δ areresistant to the p110δ-selective inhibitors PIK-39 and IC87114, butretain sensitivity to the p110α/multi-targeted inhibitors PIK-90,PIK-93, and PI-103. This chemotype-specific resistance supports theunique role of Met 752 in gating an inducible selectivity pocket.

Antagonist modeling was performed using the PyMOL Molecular GraphicsSystem. All p110γ crystal structures (PDB codes in parentheses),including the Apo (1E8Y), ATP (1E8X), Wortmannin (1E7U), LY294002(1E7V), Quercetin (1E8W), Myricetin (1E90), and Staurosporine (1E8Z),PIK-90, PIK-93, and PIK-39 bound forms were structurally aligned usingPyMOL's align function. Models for the inhibitors PIK-108, KU-55933, andPI-103 were built on top of the LY294002 arylmorpholine scaffold (1E7V)using PyMOL's fragment building function. A model for the inhibitorIC87114 was similarly built on top of the PIK-39 aryl-isoquinolinescaffold.

The model for PI-103 was built into the protein structure of p110γ boundto PIK-90, because the PIK-90 structure contains the enlarged affinitypocket that is necessary to accommodate PIK-103's phenolic moiety (thePIK-90 p110γ structure otherwise does not exhibit any conformationaldifferences in the arymorpholine-binding region in comparison to theLY294002-bound p110γ structure). The models for PIK-108, KU-55933, andIC87114 were built into the protein structure of p110γ bound to PIK-39because these inhibitors possess bulky groups that project out of theadenine plane and are likely to exploit the unique “Met 804 down”induced-fit pocket. In all inhibitor models, the choice of proteinstructure and inhibitor binding mode is based on extensive biochemicalSAR as well as inhibitor geometry. The protein structures and inhibitormodels have not been minimized to optimize binding energy, but care wastaken to prevent any gross steric clashes and to satisfy key hydrogenbonds.

C. p110α/p85α, p110β/p85α, p110δ/p85α, and p110γ Assay

The class I PI3-Ks were either purchased (p110α/p85α, p110β/p85α,p110δ/p85α from Upstate, and p110γ from Sigma) or expressed aspreviously described (Knight et al., 2004). IC50 values were measuredusing either a standard TLC assay for lipid kinase activity (describedbelow) or a high-throughput membrane capture assay. Kinase reactionswere performed by preparing a reaction mixture containing kinase,inhibitor (2% DMSO final concentration), buffer (25 mM HEPES, pH 7.4, 10mM MgCl2), and freshly sonicated phosphatidylinositol (100 μg/ml).Reactions were initiated by the addition of ATP containing 10 μCi ofγ-32P-ATP to a final concentration 10 or 100 μM, as indicated in FIG. 5,and allowed to proceed for 5 minutes at room temperature. For TLCanalysis, reactions were then terminated by the addition of 105 μl 1NHCl followed by 160 μl CHCl3:MeOH (1:1). The biphasic mixture wasvortexed, briefly centrifuged, and the organic phase transferred to anew tube using a gel loading pipette tip precoated with CHCl₃. Thisextract was spotted on TLC plates and developed for 3-4 hours in a 65:35solution of n-propanol:1M acetic acid. The TLC plates were then dried,exposed to a phosphorimager screen (Storm, Amersham), and quantitated.For each compound, kinase activity was measured at 10-12 inhibitorconcentrations representing two-fold dilutions from the highestconcentration tested (typically, 200 μM). For compounds showingsignificant activity, IC50 determinations were repeated two to fourtimes, and the reported value is the average of these independentmeasurements.

Results are set forth below in Table 2.

D. Abl Assay

Inhibitors (final concentration: 10 μM) were assayed in triplicateagainst recombinant full-length Abl or Abl (T315I) (Upstate) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl2, 200 μM ATP (2.5 μCi ofγ-32P-ATP), and 0.5 mg/mL BSA. The optimized Abl peptide substrateEAIYAAPFAKKK was used as phosphoacceptor (200 μM). Reactions wereterminated by spotting onto phosphocellulose sheets, which were washedwith 0.5% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets were dried and the transferred radioactivity quantitated byphosphorimaging.

Results are set forth below in Table 3.

E. Hck Assay

Hck: Inhibitors (final concentration: 10 μM) were assayed in triplicateagainst recombinant full-length Hck in an assay containing 25 mM HEPES,pH 7.4, 10 mM MgCl2, 200 μM ATP (2.5 μCi of γ-32P-ATP), and 0.5 mg/mLBSA. The optimized Src family kinase peptide substrate EIYGEFKKK wasused as phosphoacceptor (200 μM). Reactions were terminated by spottingonto phosphocellulose sheets, which were washed with 0.5% phosphoricacid (approximately 6 times, 5-10 minutes each). Sheets were dried andthe transferred radioactivity quantitated by phosphorimaging.

Results are set forth below in Table 3.

F. Inulsin Receptor (IR) Assay

Inhibitors (final concentration: 10 μM were assayed in triplicateagainst recombinant insulin receptor kinase domain (Upstate) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl2, 10 mM MnCl2, 200 μM ATP(2.5 μCi of γ-32P-ATP), and 0.5 mg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) wasused as a substrate. Reactions were terminated by spotting ontonitrocellulose, which was washed with 1M NaCl/1% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets were dried and thetransferred radioactivity quantitated by phosphorimaging.

Results are set forth-below in Table 4.

G. Src Assay

Src, Src (T338I): Inhibitors (final concentration: 10 μM) were assayedin triplicate against recombinant full-length Src or Src (T338I) in anassay containing 25 mM HEPES, pH 7.4, 10 mM MgCl2, 200 μM ATP (2.5 μCiof γ-32P-ATP), and 0.5 mg/mL BSA. The optimized Src family kinasepeptide substrate EIYGEFKKK was used as phosphoacceptor (200 μM).Reactions were terminated by spotting onto phosphocellulose sheets,which were washed with 0.5% phosphoric acid (approximately 6 times, 5-10minutes each). Sheets were dried and the transferred radioactivityquantitated by phosphorimaging.

Results are set forth below in Table 3.

H. DNA-PK (DNAK) Assay

DNA-PK was purchased from Promega and assayed using the DNA-PK AssaySystem (Promega) according to the manufacturer's instructions.

Results are shown in Table 2.

I. mTOR Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant mTOR (Invitrogen) in an assay containing 50 mM HEPES, pH7.5, 1 mM EGTA, 10 mM MgCl2, 2.5 mM, 0.01% Tween, 10 μM ATP (2.5 μCi ofμ-32P-ATP), and 3 μg/mL BSA. Rat recombinant PHAS-1/4EBP1 (Calbiochem; 2mg/mL) was used as a substrate. Reactions were terminated by spottingonto nitrocellulose, which was washed with 1M NaCl/1% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets were dried and thetransferred radioactivity quantitated by phosphorimaging.

Results are set forth in Table 4 below.

J. Vascular Endothelial Growth Receptor

Vascular endothelial growth receptor 2(KDR): Inhibitors (finalconcentrations 50 μM-0.003 μM) were tested against recombinant KDRreceptor kinase domain (Invitrogen) in an assay containing 25 mM HEPES,pH 7.4, 10 mM MgCl2, 0.1% BME, 10 RM ATP (2.5 μCi of μ-32P-ATP), and 3μg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) was used as a substrate. Reactionswere terminated by spotting onto nitrocellulose, which was washed with1M NaCl/1% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets were dried and the transferred radioactivity quantitated byphosphorimaging.

Results are set forth in Table 3 below.

K. Ephrin Receptor B4 (EphB4) Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant Ephrin receptor B4 kinase domain (Invitrogen) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl2, 0.1% BME, 10 μM ATP (2.5μCi of μ-32P-ATP), and 3 μg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) was usedas a substrate. Reactions were terminated by spotting ontonitrocellulose, which was washed with 1M NaCl/1% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets were dried and thetransferred radioactivity quantitated by phosphorimaging.

Results are set forth in Table 3 below.

L. Epidermal Growth Factor Receptor (EGFR) Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant EGF receptor kinase domain (Invitrogen) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl2, 0.1% BME, 10 μM ATP (2.5μCi of μ-32P-ATP), and 3 μg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) was usedas a substrate. Reactions were terminated by spotting ontonitrocellulose, which was washed with 1M NaCl/1% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets were dried and thetransferred radioactivity quantitated by phosphorimaging.

Results are set forth in Table 3 below.

M. KIT Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant KIT kinase domain (Invitrogen) in an assay containing 25 mMHEPES, pH 7.4, 10 mM MgCl2, 1 mM DTT, 10 mM MnCl2, 10 μM ATP (2.5 μCi ofμ-32P-ATP), and 3 μg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) was used as asubstrate. Reactions were terminated by spotting onto nitrocellulose,which was washed with 1M NaCl/1% phosphoric acid (approximately 6 times,5-10 minutes each). Sheets were dried and the transferred radioactivityquantitated by phosphorimaging.

Results are set forth in Table 4 below.

N. RET Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant RET kinase domain (Invitrogen) in an assay-containing 25 mMHEPES, pH 7.4, 10 mM MgCl2, 2.5 mM DTT, 10 μM ATP (2.5 μCi ofμ-32P-ATP), and 3 μg/mL BSA. The optimized Abl peptide substrateEAIYAAPFAKKK was used as phosphoacceptor (200 μM). Reactions wereterminated by spotting onto phosphocellulose sheets, which were washedwith 0.5% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets were dried and the transferred radioactivity quantitated byphosphorimaging.

Results are set forth in Table 4 below.

O. Platelet Derived Growth Factor Receptor (PDGFR) Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant PDG receptor kinase domain (Invitrogen) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl2, 2.5 mM DTT, 10 μM ATP (2.5μCi of μ-32P-ATP), and 3 μg/mL BSA. The optimized Abl peptide substrateEAIYAAPFAKKK was used as phosphoacceptor (200 μM). Reactions wereterminated by spotting onto phosphocellulose sheets, which were washedwith 0.5% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets were dried and the transferred radioactivity quantitated byphosphorimaging.

Results are set forth in Table 4 below.

P. FMS-Related Tyrosine Kinase 3 (FLT-3) Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant FLT-3 kinase domain (Invitrogen) in an assay containing 25mM HEPES, pH 7.4, 10 mM MgCl2, 2.5 mM DTT, 10 μM ATP (2.5 μCi ofμ-32P-ATP), and 3 μg/mL BSA. The optimized Abl peptide substrateEAIYAAPFAKKK was used as phosphoacceptor (200 μM). Reactions wereterminated by spotting onto phosphocellulose sheets, which were washedwith 0.5% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets were dried and the transferred radioactivity quantitated byphosphorimaging.

Results are set forth in Table 4 below.

Q. TEK Receptor Tyrosine Kinase (TIE2) Assay

Inhibitors (final concentrations 50 μM-0.003 μM) were tested againstrecombinant TIE2 kinase domain (Invitrogen) in an assay containing 25 mMHEPES, pH 7.4, 10 mM MgCl2, 2 mM DTT, 10 mM MnCl2, 10 μM ATP (2.5 μCi ofμ-32P-ATP), and 3 μg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) was used as asubstrate. Reactions were terminated by spotting onto nitrocellulose,which was washed with 1M NaCl/1% phosphoric acid (approximately 6 times,5-10 minutes each). Sheets were dried and the transferred radioactivityquantitated by phosphorimaging.

Results are set forth in Table 4 below.

R. Results

In Table 1, the compounds have the formula:

R¹ and R² are as defined in Table 1. X is ═N— except where indicated.R³⁶ is —NH₂ except where indicated. TABLE 1 Cpd R1 R2 KS167 —H

ZK141 —H

KS84

ZK127 —H

ZK134 —H

ZK132 —H

ZK125 —H

BA56

ZK138 —H

KS287

KS288

KS284

BA60

ZK318 —H

ZK320 —H

ZK333 —Me

ZK323

ZK327

BA77d

BA78d

BA22

BA79d

BA85

BAB7

ZK502

ZK489 —Me

ZK487

Zk491

Zk493

BA62d

ZK450

ZK454

ZK469

ZK471

ZK461

ZK413

ZK379

ZK421

ZK403

ZK405

ZK432

ZK434

ZK465

ZK377

ZK399

ZK401

BA62

ZK358

ZK452

ZK456

ZK463

ZK371

ZK409

ZK428

ZK430

ZK387

ZK389

ZK369

ZK385

ZK391

BA155 2

BA157 2

BA59

BA63

BA93

BA49

BA15

ZK321 —H

ZK337 —Me

ZK347

ZK325

ZK349

ZK423

ZK411

ZK407

BA98

BA23

ZK485

ZK495

ZK496

ZK494

BA90

ZK341 —H

ZK343

ZK361

ZK359

ZK362

BA64

BA65

ZK305

ZK306

BA66

BA48

ZK133 —H

BA20dd

BA20d

BA20

BA99

BA81dd

BA81d

ZK137 —H

ZK135 —H

ZK130 —H

ZK128 —H

BA26

BA46

BA45

BA39

BA150

BA151

BA21

BA52

BA53

BA31

BA152

BA149

BA32

BA55

BA35

BA34

BA38

BA40

BA41

BA14

BA12

—I BA30

ZK149 —H

ZK126 —H

ZK143 —H

ZK150 —Me X = CH

ZK136 —H

ZK131 —H

ZK151 —Me X = CH

ZK142 —H

ZK139 —H

KS207 —H

KS208 —Me

ZK102 —Me (R³⁶ is Cl)

ZK157

ZK159

ZK156

KS63

ZK158

ZK147

ZK155

ZK162

ZK165

ZK161

ZK167

ZK168

BA116 —Me

BA17

BA134

BA105

BA122

BA111 —Me

BA102

BA112

BA118

BA130

BA132

BA139

BA158

BA140

BA141

BA146

BA142

BA145

BA147

BA148 —Me

BA143

BA144

BA129

BA131

BA133

BA120 —Me

BA108

BA121

BA89

BA94

BA135

BA137

BA138

BA160

BA157 3

BA154

BA110

BA115

BA159

BA119 —Me

BA107

BA124

BA161

BA162

BA24dd

BA43

BA91

BA92

BA86

BA88

BA96

BA97

BA44

BA156

BA95

ZK129 —Me X = CH

BA54

ZK152 —Me X = CH

BA42

TABLE 2 Cpd 110α 110β 110δ 110γ DPK KS167 ++ + ++ ++ ++ ZK141 ++ ++ ++++ +++ KS84 ++ + ++ + +++ ZK127 ++ ++ ++ ++ +++ ZK134 ++ + ++ ++ +++ZK132 ++ ++ ++ ++ +++ ZK125 ++ + ++ ++ +++ BA56 +++ +++ +++ +++ +++ZK138 ++ ++ ++ ++ +++ KS287 ++ ++ ++ ++ ++ KS288 + + + +++ KS284 +++ +++++ ++ +++ BA60 ++ ++ +++ ++ +++ ZK318 ++ + ++ ++ ++ ZK320 ++ ++ +++ +++++ ZK333 +++ ++ +++ +++ +++ ZK323 +++ ++ +++ +++ +++ ZK327 +++ ++ ++++++ +++ BA77d ++ ++ +++ ++ ++ BA78d ++ ++ +++ ++ +++ BA22 +++ +++ ++++++ +++ BA79d +++ +++ +++ +++ +++ BA85 ++ ++ ++ ++ +++ BA87 +++ +++ ++++++ +++ ZK502 ++ ++ +++ ++ +++ ZK489 +++ +++ +++ +++ +++ ZK487 +++ ++++++ +++ +++ Zk491 +++ +++ +++ +++ ++ Zk493 +++ +++ +++ +++ +++ BA62d ++++++ +++ +++ ++ ZK450 ++ ++ ++ +++ + ZK454 ++ ++ ++ +++ +++ ZK469 +++ +++++ +++ +++ ZK471 +++ +++ +++ +++ ++ ZK461 +++ ++ +++ ++ ++ ZK413 ++ ++++++ +++ +++ ZK379 +++ ++ +++ +++ +++ ZK421 +++ ++ +++ +++ +++ ZK403 +++++ +++ +++ ++ ZK405 ++ ++ +++ ++ +++ ZK432 +++ ++ +++ +++ +++ ZK434 ++++++ +++ +++ +++ ZK465 +++ + +++ +++ +++ ZK377 ++ ++ ++ ++ +++ ZK399 ++++ +++ ++ +++ ZK401 ++ + ++ + +++ BA62 +++ ++ +++ +++ +++ ZK358 +++ +++++ +++ +++ ZK452 ++ ++ ++ +++ +++ ZK456 +++ ++ ++ +++ +++ ZK463 +++ +++++ +++ +++ ZK371 +++ +++ +++ +++ +++ ZK409 +++ +++ +++ +++ +++ ZK428+++ +++ +++ +++ +++ ZK430 +++ +++ +++ +++ +++ ZK387 ++ ++ ++ ++ +++ZK389 ++ +++ +++ ++ +++ ZK369 ++ + ++ ++ ++ ZK385 ++ ++ +++ ++ +++ ZK391++ ++ +++ ++ +++ BA155_2 +++ +++ +++ +++ +++ BA157_2 +++ +++ +++ +++ +++BA59 +++ ++ +++ +++ +++ BA63 ++ +++ +++ BA93 +++ ++ +++ +++ +++ BA49++ + ++ ++ ++ BA15 +++ ++ +++ +++ +++ ZK321 +++ ++ +++ +++ +++ ZK337 ++++++ +++ +++ +++ ZK347 +++ +++ +++ +++ +++ ZK325 +++ +++ +++ +++ +++ZK349 +++ +++ +++ +++ +++ ZK423 +++ +++ +++ +++ +++ ZK411 ++ +++ +++ ++++++ ZK407 ++ +++ +++ ++ ++ BA98 +++ +++ +++ +++ +++ S1 ++ + ++ + + BA23+++ +++ +++ +++ +++ ZK485 +++ +++ +++ ++ ++ ZK495 +++ ++ +++ ++ +++ZK496 ++ ++ +++ ++ +++ ZK494 +++ +++ +++ +++ +++ BA90 +++ +++ +++ ++++++ ZK341 ++ ++ +++ +++ ++ ZK343 +++ ++ +++ ++ +++ ZK361 ++ ++ +++ ++++++ ZK359 +++ +++ +++ +++ +++ ZK362 +++ +++ +++ +++ ++ BA64 + + ++ + ++BA65 + + ++ + ++ ZK305 +++ +++ +++ +++ +++ ZK306 ++ + ++ ++ ++ BA66 ++++ +++ ++ +++ BA48 ++ ++ +++ ++ +++ ZK133 + + + ++ ++ BA20dd ++ ++ ++ +++++ BA20d ++ ++ +++ +++ +++ BA20 ++ ++ ++ ++ BA99 ++ + ++ + ++ BA81dd+++ ++ +++ +++ + BA81d +++ ++ +++ +++ +++ ZK137 ++ + ++ ++ +++ ZK135++ + + ++ ++ ZK130 + + + + + ZK128 ++ + ++ ++ ++ BA26 ++ ++ +++ +++ ++SU11248 ++ + ++ ++ + BAY43- ++ + + + + 9006 Dasatinib ++ + ++ + ++ BA46++ ++ +++ ++ +++ BA45 ++ + ++ ++ ++ BA39 ++ +++ ++ ++ BA150 ++ + ++ ++++ BA151 ++ + ++ ++ ++ BA21 ++ ++ ++ ++ BA52 ++ ++ ++ ++ ++ BA53++ + + + ++ BA31 ++ ++ +++ +++ ++ BA152 ++ ++ +++ ++ +++ BA149 ++ ++ ++++ + BA32 ++ ++ +++ +++ BA55 ++ ++ +++ ++ ++ BA35 ++ +++ +++ +++ BA34 ++++ +++ ++ +++ BA38 ++ +++ +++ +++ BA40 ++ ++ ++ +++ BA41 ++ ++ +++ ++++++ BA14 ++ ++ +++ ++ +++ BA12 ++ ++ +++ ++ ++ BA30 ++ ++ +++ +++ +++ZK149 + ++ ++ ++ + ZK126 ++ + ++ + ++ ZK143 + + + + + ZK150 ++ + + + ++ZK136 ++ + + ++ +++ ZK131 + + + + + ZK151 + + + + +++ ZK142 + + + + +ZK139 + + + ++ ++ KS207 ++ + +++ +++ +++ KS208 ++ ++ ++ +++ +++ZK102 + + + + +++ ZK157 +++ +++ ZK159 ++ ++ ++ ++ +++ ZK156 ++ ++ ++ +++++ KS63 + + + + +++ ZK158 ++ ++ ++ ++ +++ ZK147 + ++ ++ ++ ++ ZK155 ++++ ++ ++ +++ ZK162 ++ + ++ ++ +++ ZK165 ++ + ++ ++ +++ ZK161 ++ + ++ ++++ ZK167 ++ ++ ZK168 ++ +++ BA116 + ++ ++ ++ +++ BA17 +++ ++ +++ ++ +++BA134 ++ + ++ + ++ BA105 ++ ++ +++ ++ +++ BA122 ++ ++ +++ ++ +++ BA111+++ ++ +++ +++ +++ BA102 +++ +++ +++ +++ +++ BA112 +++ ++ +++ +++ +++BA118 +++ ++ +++ +++ +++ BA130 +++ ++ +++ +++ +++ BA132 +++ +++ +++ ++++++ BA139 ++ ++ +++ + ++ BA158 +++ +++ +++ +++ +++ BA140 + ++ + + ++BA141 + + + + + BA146 +++ +++ +++ +++ +++ BA142 +++ ++ ++ +++ +++ BA145++ ++ ++ ++ ++ BA147 ++ ++ ++ ++ ++ BA148 ++ ++ ++ ++ +++ BA143 ++ ++ ++++ +++ BA144 +++ ++ +++ +++ +++ BA129 ++ + ++ + ++ BA131 +++ ++ +++ ++++++ BA133 +++ +++ +++ +++ +++ BA120 +++ ++ +++ +++ +++ BA108 +++ ++ +++++ +++ BA121 +++ ++ +++ ++ +++ BA89 +++ ++ +++ +++ +++ BA94 +++ ++ ++++++ +++ BA135 +++ +++ +++ +++ +++ BA136 +++ ++ +++ ++ +++ BA137 +++ ++++++ +++ +++ BA138 +++ ++ +++ +++ +++ BA160 ++ ++ +++ +++ +++ BA157_3 +++++ +++ +++ +++ BA154 +++ +++ +++ +++ +++ BA110 +++ ++ +++ +++ +++ BA115+++ ++ +++ +++ +++ BA159 +++ +++ +++ +++ ++ BA119 ++ ++ +++ ++ +++ BA107++ ++ +++ ++ +++ BA124 ++ ++ +++ ++ +++ BA161 + + + + +++ BA162 ++ ++ ++++ +++ BA24dd ++ ++ +++ ++ +++ BA43 +++ +++ +++ +++ BA91 ++ ++ ++ ++ +++BA92 ++ ++ +++ ++ +++ BA86 ++ ++ ++ +++ +++ BA88 + + ++ +++ +++ BA96++ + +++ +++ +++ BA97 ++ ++ +++ +++ +++ BA44 ++ +++ ++ ++ ++ BA156 ++ ++++ ++ +++ BA95 ++ ++ +++ +++ +++ ZK129 ++ ++ ++ ++ +++ BA54 ++ ++ ++ +++++ ZK152 ++ ++ ++ ++ +++ BA42 ++ ++ ++ ++ +++ PIK294 ++ +++ +++ +++ ++Iressa + + + + + PIK103 +++ +++ +++ +++ +++ PIK90 +++ +++ +++ +++ +++

TABLE 3 Cpd Abl Hck Src Src (T/I) VEGFR EGFR EphB4 KS84 +++ +++ +++ +++++ +++ +++ BA56 ++ +++ +++ ++ ++ +++ ++ KS284 +++ +++ +++ ++ +++ ++++++ BA60 ++ +++ +++ +++ ++ + ++ ZK318 ++ ++ +++ ++ + ++ ZK320 +++ ++++++ ++ +++ ++ ++ ZK333 +++ +++ +++ ++ +++ ++ ++ ZK323 +++ +++ +++ ++++++ +++ +++ ZK327 +++ +++ +++ ++ +++ ++ ++ BA77d +++ +++ +++ ++ ++ +++++ BA78d +++ +++ +++ ++ +++ ++ ++ BA22 +++ +++ +++ ++ +++ +++ +++ BA79d+++ +++ +++ ++ +++ +++ +++ BA85 ++ +++ +++ ++ ++ ++ ++ BA87 +++ +++ ++++++ +++ +++ +++ ZK502 +++ +++ +++ ++ ++ ++ + ZK489 +++ +++ +++ + ++ + ++ZK487 +++ +++ +++ ++ +++ +++ +++ Zk491 +++ +++ +++ ++ +++ +++ +++ Zk493+++ +++ +++ ++ +++ +++ +++ BA62d +++ +++ +++ +++ +++ ++ +++ ZK450 ++++++ +++ + ++ + ++ ZK454 +++ ++ +++ + ++ ++ + ZK469 +++ +++ +++ + ++ ++++ ZK471 +++ +++ +++ + ++ ++ + ZK461 +++ +++ +++ + +++ ++ ++ ZK413 ++++++ +++ + ++ ++ ZK379 +++ +++ +++ ++ +++ +++ +++ ZK421 +++ +++ +++ ++++++ +++ +++ ZK403 +++ +++ +++ ++ +++ ++ +++ ZK405 +++ +++ +++ ++ ++ ++ +ZK432 +++ +++ +++ ++ +++ ++ +++ ZK434 +++ +++ +++ ++ +++ ++ +++ ZK465+++ +++ +++ + +++ +++ +++ ZK377 ++ ++ +++ + ++ + + ZK399 +++ ++++++ + + + ++ ZK401 +++ +++ +++ + + + + BA62 +++ +++ +++ + + ++ +++ ZK358+++ +++ +++ ++ +++ +++ +++ ZK452 +++ +++ +++ ++ +++ ++ ++ ZK456 +++ ++++++ + ++ ++ ++ ZK463 +++ +++ +++ ++ +++ ++ ++ ZK371 +++ +++ +++ ++ ++++++ +++ ZK409 +++ +++ +++ ++ +++ ++ +++ ZK428 +++ +++ +++ ++ +++ ++ +++ZK430 +++ +++ +++ ++ +++ ++ +++ ZK387 +++ +++ +++ + ++ ++ ++ ZK389 ++++++ +++ ++ +++ ++ +++ ZK369 ++ ++ +++ + ++ ++ + ZK385 ++ +++ +++ + ++ +++ ZK391 ++ +++ +++ + ++ ++ ++ BA155_2 ++ +++ +++ ++ ++ ++ ++ BA157_2 ++++ ++ ++ ++ ++ ++ BA59 +++ +++ +++ +++ ++ ++ +++ BA63 ++ +++ +++ ++ ++++ ++ BA93 ++ +++ +++ ++ ++ ++ ++ BA49 ++ +++ +++ ++ + ++ +++ BA15 +++++ +++ ++ + +++ ++ ZK321 +++ +++ +++ ++ ++ + ++ ZK337 +++ +++ +++ + ++ZK347 +++ +++ +++ ++ +++ ++ ++ ZK325 +++ +++ +++ ++ +++ ++ +++ ZK349 ++++++ +++ +++ +++ +++ +++ ZK423 +++ +++ +++ ++ +++ +++ +++ ZK411 ++ +++++ + ++ ++ ++ ZK407 ++ +++ +++ + +++ ++ + BA98 +++ +++ +++ ++ +++ +++++ S1 +++ + ++ + + + + BA23 ++ ++ +++ ++ + ++ ++ ZK485 ++ +++ +++ + + ++++ ZK495 ++ +++ ++ ++ + ++ ++ ZK496 ++ ++ ++ ++ ++ ++ ++ ZK494 +++ ++++++ ++ ++ ++ ++ BA90 ++ +++ +++ ++ ++ ++ ++ ZK341 ++ ++ + + + + + ZK343++ +++ +++ + ++ ++ ++ ZK361 ++ ++ ++ + + ++ + ZK359 +++ +++ +++ ++ ++ +++++ ZK362 ++ +++ +++ + + ++ + BA64 +++ +++ +++ ++ + ++ +++ BA65 ++ ++++++ ++ ++ ++ ++ ZK305 ++ ++ ++ ++ + + ++ ZK306 + ++ ++ ++ ++ ++ + BA66++ ++ ++ + + ++ BA48 ++ +++ +++ ++ ++ ++ ++ BA20dd +++ +++ +++ ++ ++ ++++ BA20d +++ +++ +++ ++ ++ ++ ++ BA20 +++ +++ +++ +++ ++ +++ + BA99 +++++ +++ ++ +++ ++ ++ BA81dd +++ +++ +++ ++ +++ +++ +++ BA81d +++ +++ +++++ +++ +++ +++ BA26 ++ ++ +++ ++ + + + BA46 ++ ++ ++ + ++ ++ ++ BA45 +++++ +++ ++ ++ ++ + BA39 ++ ++ ++ ++ ++ + ++ BA150 ++ ++ ++ + ++ + ++BA151 ++ ++ ++ + + ++ ++ BA21 ++ ++ ++ + + + + BA52 ++ ++ ++ + + + +BA53 ++ ++ ++ + + + + BA31 ++ ++ ++ + + ++ + BA152 +++ +++ +++ + ++ ++++ BA149 ++ +++ +++ ++ ++ ++ ++ BA32 ++ ++ ++ + + + ++ BA55 ++ ++ ++ + +++ ++ BA35 ++ ++ ++ ++ ++ ++ ++ BA34 ++ ++ ++ ++ ++ ++ ++ BA38 ++ ++ ++++ + + + BA40 ++ ++ ++ ++ ++ ++ + BA41 ++ ++ ++ ++ ++ + ++ BA14 ++ ++ ++++ + + ++ BA12 ++ ++ ++ ++ ++ ++ ++ BA30 ++ ++ ++ ++ ++ +++ ++ KS208 +++++ ++ ++ ++ +++ ++ BA116 ++ +++ ++ +++ ++ ++ BA17 +++ +++ +++ +++ ++++++ +++ BA134 ++ ++ ++ ++ + + ++ BA105 ++ +++ ++ ++ ++ ++ ++ BA122 ++++++ +++ ++ ++ +++ +++ BA111 + + ++ ++ ++ ++ ++ BA102 +++ +++ +++ ++ ++++++ +++ BA112 ++ +++ +++ ++ ++ ++ ++ BA118 ++ ++ + ++ ++ + ++ BA130 +++++ ++ ++ +++ + ++ BA132 ++ +++ ++ + ++ ++ ++ BA139 ++ ++ ++ + ++ ++ +BA158 +++ +++ +++ ++ ++ ++ ++ BA140 ++ ++ ++ ++ + ++ ++ BA141 + ++ + + +++ + BA146 ++ ++ ++ + ++ ++ ++ BA142 ++ ++ +++ + ++ ++ ++ BA145 + ++++ + + + ++ BA147 ++ ++ ++ + ++ + ++ BA148 + ++ ++ + + ++ + BA143 ++ ++++ ++ + ++ ++ BA144 ++ ++ +++ + ++ + ++ BA129 ++ + +++ + ++ ++ ++ BA131++ ++ 11 + ++ + ++ BA133 ++ ++ +++ + + ++ ++ BA120 +++ +++ +++ ++ +++ ++++ BA108 +++ +++ +++ +++ +++ +++ +++ BA121 +++ +++ +++ +++ +++ +++ +++BA89 +++ +++ +++ +++ +++ +++ +++ BA94 +++ +++ +++ +++ +++ +++ +++ BA135+++ +++ +++ ++ +++ +++ + BA136 +++ +++ +++ ++ +++ ++ BA137 +++ +++ ++++++ +++ ++ ++ BA138 +++ +++ +++ +++ +++ +++ ++ BA160 +++ +++ +++ ++ +++++ ++ BA157_3 ++ +++ +++ ++ + ++ ++ BA154 +++ +++ +++ ++ ++ ++ ++ BA110+++ +++ +++ ++ +++ ++ ++ BA115 +++ +++ +++ +++ +++ +++ BA159 +++ +++ +++++ +++ ++ ++ BA119 ++ +++ +++ ++ ++ ++ ++ BA107 +++ +++ +++ ++ +++ ++++++ BA124 +++ +++ +++ +++ +++ +++ +++ BA161 ++ + ++ + + ++ + BA162 ++ ++++ + + ++ ++ BA24dd ++ ++ ++ ++ ++ ++ ++ BA43 ++ +++ ++ ++ ++ ++ ++ BA91++ ++ +++ ++ ++ ++ ++ BA92 ++ +++ +++ ++ ++ ++ ++ BA86 +++ +++ +++ ++++++ +++ +++ BA88 +++ +++ +++ +++ +++ +++ +++ BA96 +++ +++ +++ ++ ++ ++++++ BA97 +++ +++ +++ ++ ++ +++ +++ BA44 ++ ++ ++ ++ + ++ + BA156 ++ ++++ ++ ++ +++ ++ BA95 ++ +++ ++ + + ++ ++ BA54 ++ ++ +++ ++ ++ ++ ++BA42 + ++ ++ ++ ++ + ++ SU11248 +++ +++ +++ +++ +++ ++ + BAY43- +++ ++++++ +++ +++ + ++ 9006 Dasatinib ++ + +++ Iressa ++ +++ +++ ++ ++ ++PIK103 + + + + + + + PIK90 + + + + + + + PIK294 ++ + ++ + + + +

TABLE 4 Cpd cKIT Tie2 FLT3 PDGFR RET IR mTOR ZK358 +++ ++ +++ +++ +++ +++++ ZK487 +++ +++ +++ +++ +++ + +++ ZK349 +++ +++ +++ +++ +++ ++ +++ZK494 +++ ++ ++ +++ +++ + +++ BA102 +++ +++ +++ +++ +++ + +++ BA121 ++++++ +++ +++ ++ +++ KS84 +++ + +++ +++ +++ ++ ++ SU11248 +++ ++ +++ ++++++ ++ + BAY43-9006 +++ +++ + +++ +++ ++ + Dasatinib +++ ++ +++ +++++ + + Iressa + ++ +++ +++ ++ + +In Tables 2-4 above, a +++ indicates an IC50 of less than 1 μM; a ++indicates an IC50 of from 1 μM to 50 μM; and a + indicates and IC50 ofmore than 50 μM.

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1. A compound having the formula:

wherein X is ═N— or ═C(H)—; R¹ is hydrogen, R³-substituted orunsubstituted alkyl, R³-substituted or unsubstituted heteroalkyl,R³-substituted or unsubstituted cycloalkyl, R³-substituted orunsubstituted heterocycloalkyl, R³-substituted or unsubstituted aryl, orR³-substituted or unsubstituted heteroaryl; R² is halogen,R⁴-substituted aryl, or substituted or unsubstituted heteroaryl; R³ ishalogen, —CN, —OR⁵, —S(O)_(n)R⁶, —NR⁷R⁸, —C(O)R⁹, ═N—NH₂, —NR¹⁰—C(O)R¹¹,—NR¹²—C(O)—OR¹³, —C(O)NR¹⁴R¹⁵, —NR¹⁶S(O)₂R¹⁷, —S(O)₂NR¹⁸,R¹⁹-substituted or unsubstituted alkyl, R¹⁹-substituted or unsubstitutedheteroalkyl, R¹⁹-substituted or unsubstituted cycloalkyl,R¹⁹-substituted or unsubstituted heterocycloalkyl, R¹⁹-substituted orunsubstituted aryl, or R¹⁹-substituted or unsubstituted heteroaryl,wherein n is an integer from 0 to 2; R³⁶ is —NR³⁷R³⁸ or halogen; R⁴ ishalogen, —CN, —OR²⁰, —S(O)_(q)R²¹, —NR²²R²³, —C(O)R²⁴, ═N—NH₂,—NR²⁵—C(O)R²⁶, —NR²⁷—C(O)—OR²⁸, —C(O)NR²⁹R³⁰, —NR³¹S(O)₂R³², —S(O)₂NR³³,R³⁴-substituted or unsubstituted alkyl, R³⁴-substituted or unsubstitutedheteroalkyl, R³⁴-substituted or unsubstituted cycloalkyl,R³⁴-substituted or unsubstituted heterocycloalkyl, R³⁴-substituted orunsubstituted aryl, or R³⁴-substituted or unsubstituted heteroaryl,wherein q is an integer from 0 to 2; R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹²,R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷,R²⁸, R²⁹, R³⁰, R³¹, R³², and R³³ are independently hydrogen,R³⁵-substituted or unsubstituted alkyl, R³⁵-substituted or unsubstitutedheteroalkyl, unsubstituted cycloalkyl, R³⁵-substituted or unsubstitutedheterocycloalkyl, R³⁵-substituted or unsubstituted aryl, orR³⁵-substituted or unsubstituted heteroaryl; R¹⁹, R³⁴ and R³⁵ areindependently hydrogen, halogen, unsubstituted alkyl, unsubstitutedheteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,unsubstituted aryl, or unsubstituted heteroaryl; and R³⁷and R³⁸ arehydrogen, halogen, or unsubstituted alkyl.
 2. The compound of claim 1,wherein R² is

wherein W¹, W², W³, and W⁴ are independently ═CH—, ═CR⁴—, or ═N—; andring A is a partially or fully unsaturated 6- or 7-membered ring.
 3. Thecompound of claim 1, wherein R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³,R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸,R²⁹, R³⁰, R³¹, R³², and R³³ are independently hydrogen, unsubstitutedalkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl,unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstitutedheteroaryl.
 4. The compound of claim 1, wherein R²⁰, R²¹, R²², R²³, R²⁴,R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², and R³³ are independentlyhydrogen, unsubstituted alkyl, or unsubstituted heteroalkyl.
 5. Thecompound of claim 1, wherein R¹ is R³-substituted or unsubstitutedalkyl, R³-substituted or unsubstituted cycloalkyl, or R³-substituted orunsubstituted aryl.
 6. The compound of claim 1, wherein R¹ isR³-substituted or unsubstituted alkyl, or R³-substituted orunsubstituted cycloalkyl.
 7. The compound of claim 1, wherein R¹ isR³-substituted or unsubstituted C₁-C₄ alkyl, or R³-substituted orunsubstituted C₃-C₆ cycloalkyl.
 8. The compound of claim 1, wherein R¹is R³-substituted or unsubstituted C₁-C₄ alkyl, or R³-substituted orunsubstituted cyclopentyl.
 9. The compound of claim 1, wherein R¹ ismethyl or unsubstituted C₃-C₆ branched alkyl.
 10. The compound of claim1, wherein R¹ is isopropyl.
 11. The compound of claim 1, wherein R³ isR¹⁹-substituted or unsubstituted alkyl, R¹⁹-substituted or unsubstitutedcycloalkyl, or R¹⁹-substituted or unsubstituted aryl.
 12. The compoundof claim 1, wherein R³ is R¹⁹-substituted or unsubstituted alkyl,R¹⁹-substituted or unsubstituted cycloalkyl, or R¹⁹-substituted orunsubstituted aryl.
 13. The compound of claim 1, wherein R³ isR¹⁹-substituted or unsubstituted alkyl, or R¹⁹-substituted orunsubstituted cycloalkyl.
 14. The compound of claim 1, wherein R¹⁹ isunsubstituted alkyl or unsubstituted cycloalkyl.
 15. The compound ofclaim 1, wherein R¹⁹ is unsubstituted C₁-C₄ alkyl or unsubstitutedcyclopentyl.
 16. The compound of claim 1, wherein R² is R⁴-substitutedaryl, or R⁴-substituted or unsubstituted heteroaryl.
 17. The compound ofclaim 1, wherein R² is R⁴-substituted phenyl, R⁴-substituted orunsubstituted naphthyl, R⁴-substituted or unsubstituted pyridinyl,R⁴-substituted or unsubstituted pyrimidinyl, R⁴-substituted orunsubstituted thiophenyl, R⁴-substituted or unsubstituted furanyl,R⁴-substituted or unsubstituted indolyl, R⁴-substituted or unsubstitutedbenzoxadiazolyl, R⁴-substituted or unsubstituted benzodioxolyl,R⁴-substituted or unsubstituted benzodioxanyl, R⁴-substituted orunsubstituted thianaphthanyl, R⁴-substituted or unsubstitutedpyrrolopyridinyl, R⁴-substituted or unsubstituted indazolyl,R⁴-substituted or unsubstituted tetrahydronaphthalenyl, R⁴-substitutedor unsubstituted quinolinyl, R⁴-substituted or unsubstitutedquinoxalinyl, R⁴-substituted or unsubstituted pyridopyrazinyl,R⁴-substituted or unsubstituted quinazolinonyl, R⁴-substituted orunsubstituted chromenonyl, R⁴-substituted or unsubstitutedbenzoisoxazolyl, R⁴-substituted or unsubstituted imidazopyridinyl,R⁴-substituted or unsubstituted benzofuranyl, R⁴-substituted orunsubstituted dihydro-benzofuranyl, R⁴-substituted or unsubstituteddihydro-benzodioxinyl, R⁴-substituted or unsubstitutedbenzoimidazolonyl, or R⁴-substituted or unsubstituted benzothiophenyl.18. The compound of claim 1, wherein R² is R⁴-substituted phenyl,R⁴-substituted or unsubstituted pyrrolpyridinyl, R⁴-substituted orunsubstituted quinolinyl, R⁴-substituted or unsubstituted indazolyl,R⁴-substituted or unsubstituted quinolinyl indolyl, or R⁴-substituted orunsubstituted naphthyl.
 19. The compound of claim 18, wherein R⁴ ishalogen, —CN, —OR²⁰, or —NR²²R²³.
 20. The compound of claim 18, whereinR⁴ is halogen, or —OR²⁰.
 21. The compound of claim 20, wherein R²⁰ ishydrogen or unsubstituted C₁-C₁₀ alkyl.
 22. The compound of claim 20,wherein R²⁰ is hydrogen or unsubstituted C₁-C₄ alkyl.
 23. The compoundof claim 20, wherein R²⁰ is hydrogen or methyl.
 24. The compound ofclaim 1, wherein R² has the formula:

R⁴ is absent, halogen, unsubstituted C₁-C₄ alkyl, or —OR²⁰, wherein R²⁰is unsubstituted C₁-C₄ alkyl.
 25. The compound of claim 1, wherein R² is6-hydroxynaphthyl, unsubstituted 7-azaindole, unsubstituted indolyl,unsubstituted indazolyl, or unsubstituted quinolinyl.
 26. The compoundof claim 1, wherein X is ═N—.
 27. The compound of claim 1, wherein R³⁶is —NH₂.
 28. A pharmaceutical composition comprising the compound ofclaim 1 and a pharmaceutically acceptable excipient.
 29. A method ofdecreasing the catalytic activity of a PI3 kinase, the method comprisingthe step of contacting said PI3 kinase with an activity decreasingamount of an antagonist, wherein said antagonist is a PI3-Kinaseaffinity pocket binding antagonist.
 30. The method of claim 29, whereinsaid antagonist is a PI3-Kinase affinity pocket bindingpyrazolopyrimidine antagonist or a PI3-Kinase affinity pocket bindingpyrrolopyrimidine antagonist.
 31. The method of claim 29, wherein saidantagonist is capable of decreasing the catalytic activity of a tyrosinekinase.
 32. The method of claim 29, wherein said antagonist is capableof decreasing the catalytic activity of a mTOR.
 33. The method of claim29, wherein said antagonist is capable of decreasing the catalyticactivity of a mTOR and a tyrosine kinase.
 34. The method of claim 29,wherein said PI3 kinase is p110α.
 35. A method of decreasing thecatalytic activity of a PI3 kinase, the method comprising the step ofcontacting said PI3 kinase with an activity decreasing amount of anantagonist, wherein said antagonist the formula:

wherein X is ═N— or ═C(H)—; R¹ is hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl; and R² is halogen, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl; R³⁶is halogen, —NR³⁷R³⁸, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl; and R³⁷and R³⁸ are independently hydrogen, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl. 36.The method of claim 35, wherein: R¹ is hydrogen, R³-substituted orunsubstituted alkyl, R³-substituted or unsubstituted heteroalkyl,R³-substituted or unsubstituted cycloalkyl, R³-substituted orunsubstituted heterocycloalkyl, R³-substituted or unsubstituted aryl, orR³-substituted or unsubstituted heteroaryl; R² is halogen,R⁴-substituted aryl, or substituted or unsubstituted heteroaryl; R³ ishalogen, —CN, —OR⁵, —S(O)_(n)R⁶, —NR⁷R⁸, —C(O)R⁹, ═N—NH₂, —NR¹⁰—C(O)R¹¹,—NR¹²—C(O)—OR¹³, —C(O)NR¹⁴R¹⁵, —NR¹⁶S(O)₂R¹⁷, —S(O)₂NR¹⁸,R¹⁹-substituted or unsubstituted alkyl, R¹⁹-substituted or unsubstitutedheteroalkyl, R¹⁹-substituted or unsubstituted cycloalkyl,R¹⁹-substituted or unsubstituted heterocycloalkyl, R¹⁹-substituted orunsubstituted aryl, or R¹⁹-substituted or unsubstituted heteroaryl,wherein n is an integer from 0 to 2; R³⁶ is NR³⁷R³⁸ or halogen; R⁴ ishalogen, —CN, —OR²⁰, —S(O)_(q)R²¹, —NR²²R²³, —C(O)R²⁴, ═N—NH₂,—NR²⁵—C(O)R²⁶, —NR²⁷—C(O)—OR²⁸, —C(O)NR²⁹R³⁰, —NR³¹S(O)₂R³², —S(O)₂NR³³,R³⁴-substituted or unsubstituted alkyl, R³⁴-substituted or unsubstitutedheteroalkyl, R³⁴-substituted or unsubstituted cycloalkyl,R³⁴-substituted or unsubstituted heterocycloalkyl, R³⁴-substituted orunsubstituted aryl, or R³⁴-substituted or unsubstituted heteroaryl,wherein q is an integer from 0 to 2; R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹²,R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷,R²⁸, R²⁹, R³⁰, R³¹, R³², and R³³ are independently hydrogen,R³⁵-substituted or unsubstituted alkyl, R³⁵-substituted or unsubstitutedheteroalkyl, unsubstituted cycloalkyl, R³⁵-substituted or unsubstitutedheterocycloalkyl, R³⁵-substituted or unsubstituted aryl, orR³⁵-substituted or unsubstituted heteroaryl; R¹⁹, R³⁴ and R³⁵ areindependently hydrogen, halogen, unsubstituted alkyl, unsubstitutedheteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,unsubstituted aryl, or unsubstituted heteroaryl; and R³⁷ and R³⁸ arehydrogen, halogen, or unsubstituted alkyl.
 37. The method of claim 35,wherein R³⁶ is —NH₂.
 38. The method of claim 35, wherein R² has theformula:

wherein W¹, W², W³, and W⁴ are independently ═CH—, ═CR⁴—, or ═N—; andring A is a partially or fully unsaturated 6- or 7-membered ring.
 39. Amethod of treating a disease mediated by PI3 kinase activity, or PI3Kinase activity and Tyrosine Kinase Activity, in a subject in need ofsuch treatment, said method comprising administering to said subject atherapeutically effective amount of an antagonist, wherein saidantagonist is a PI3-Kinase affinity pocket binding pyrazolopyrimidineantagonist or a PI3-Kinase affinity pocket binding pyrrolopyrimidineantagonist.
 40. The method of claim 39, wherein said disease is livercancer, colon cancer, breast cancer, melanoma, acute myelogenousleukemia, chronic myelogenous leukemia, or nonsmall-cell lung cancer.41. A method of treating a disease mediated by PI3 kinase activity, orPI3 Kinase activity and Tyrosine Kinase Activity, in a subject in needof such treatment, said method comprising administering to said subjecta therapeutically effective amount of an antagonist, wherein saidantagonist has the formula:

wherein X is ═N— or ═C(H)—; R¹ is hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl; and R² is halogen, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl; R³⁶is halogen, —NR³⁷R³⁸, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl; and R³⁷and R³⁸ are independently hydrogen, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl. 42.The method of claim 41, wherein: R¹ is hydrogen, R³-substituted orunsubstituted alkyl, R³-substituted or unsubstituted heteroalkyl,R³-substituted or unsubstituted cycloalkyl, R³-substituted orunsubstituted heterocycloalkyl, R³-substituted or unsubstituted aryl, orR³-substituted or unsubstituted heteroaryl; R² is halogen,R⁴-substituted aryl, or substituted or unsubstituted heteroaryl; R³ ishalogen, —CN, —OR⁵, —S(O)_(n)R⁶, —NR⁷R⁸, —C(O)R⁹, ═N—NH₂, —NR¹⁰—C(O)R¹¹,—NR¹²—C(O)—OR¹³, —C(O)NR¹⁴R¹⁵, —NR¹⁶S(O)₂R¹⁷, —S(O)₂NR¹⁸,R¹⁹-substituted or unsubstituted alkyl, R¹⁹-substituted or unsubstitutedheteroalkyl, R¹⁹-substituted or unsubstituted cycloalkyl,R¹⁹-substituted or unsubstituted heterocycloalkyl, R¹⁹-substituted orunsubstituted aryl, or R¹⁹-substituted or unsubstituted heteroaryl,wherein n is an integer from 0 to 2; R³⁶ is NR³⁷R³⁸ or halogen; R⁴ ishalogen, —CN, —OR²⁰, —S(O)_(q)R²¹, —NR²²R²³, —C(O)R²⁴, ═N—NH₂,—NR²⁵—C(O)R²⁶, —NR²⁷—C(O)—OR²⁸, —C(O)NR²⁹R³⁰, —NR³¹S(O)₂R³², —S(O)₂NR³³,R³⁴-substituted or unsubstituted alkyl, R³⁴-substituted or unsubstitutedheteroalkyl, R³⁴-substituted or unsubstituted cycloalkyl,R³⁴-substituted or unsubstituted heterocycloalkyl, R³⁴-substituted orunsubstituted aryl, or R³⁴-substituted or unsubstituted heteroaryl,wherein q is an integer from 0 to 2; R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹²,R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷,R²⁸, R²⁹, R³⁰, R³¹, R³², and R³³ are independently hydrogen,R³⁵-substituted or unsubstituted alkyl, R³⁵-substituted or unsubstitutedheteroalkyl, unsubstituted cycloalkyl, R³⁵-substituted or unsubstitutedheterocycloalkyl, R³⁵-substituted or unsubstituted aryl, orR³⁵-substituted or unsubstituted heteroaryl; R¹⁹, R³⁴ and R³⁵ areindependently hydrogen, halogen, unsubstituted alkyl, unsubstitutedheteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,unsubstituted aryl, or unsubstituted heteroaryl; and R³⁷and R³⁸ arehydrogen, halogen, or unsubstituted alkyl.
 43. The method of claim 41,wherein R³⁶ is —NH₂.
 44. The method of claim 41, wherein R² has theformula:

wherein W¹, W², W³, and W⁴ are independently ═CH—, ═CR⁴—, or ═N—; andring A is a partially or fully unsaturated 6- or 7-membered ring. 45.The method of claim 40, wherein said disease is liver cancer, coloncancer, breast cancer, melanoma, acute myelogenous leukemia, chronicmyelogenous leukemia, or nonsmall-cell lung cancer.
 46. A method ofdisrupting the function of a leukocyte or disrupting a function of anosteoclast, said method comprising contacting said leukocyte or saidosteoclast with a function disrupting amount of an antagonist, whereinsaid antagonist is a PI3-Kinase affinity pocket bindingpyrazolopyrimidine antagonist or a PI3-Kinase affinity pocket bindingpyrrolopyrimidine antagonist.